Polycyclic-carbamoylpyridone compounds and their pharmaceutical use

ABSTRACT

Compounds for use in the treatment of human immunodeficiency virus (HIV) infection are disclosed. The compounds have the following Formula (I): 
     
       
         
         
             
             
         
       
     
     including stereoisomers and pharmaceutically acceptable salts thereof, wherein R 1 , X, W, Y 1 , Y 2 , Z 1 , and Z 4  are as defined herein. Methods associated with preparation and use of such compounds, as well as pharmaceutical compositions comprising such compounds, are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application claims the benefit under 35 U.S.C. §119(e) ofU.S. Provisional Patent Application No. 61/745,375, filed Dec. 21, 2012,U.S. Provisional Patent Application No. 61/788,397, filed Mar. 15, 2013,and U.S. Provisional Patent Application No. 61/845,803, filed Jul. 12,2013. The foregoing applications are incorporated herein by reference intheir entireties.

BACKGROUND

Field

Compounds, compositions, and methods for the treatment of humanimmunodeficiency virus (HIV) infection are disclosed. In particular,novel polycyclic carbamoylpyridone compounds and methods for theirpreparation and use as therapeutic or prophylactic agents are disclosed.

Description of the Related Art

Human immunodeficiency virus infection and related diseases are a majorpublic health problem worldwide. Human immunodeficiency virus type 1(HIV-1) encodes three enzymes which are required for viral replication:reverse transcriptase, protease, and integrase. Although drugs targetingreverse transcriptase and protease are in wide use and have showneffectiveness, particularly when employed in combination, toxicity anddevelopment of resistant strains have limited their usefulness (Palella,et al. N. Fang. J Med. (1998) 338:853-860; Richman, D. D. Nature (2001)410:995-1001).

Pregnane X receptor (PXR) is a nuclear receptor that is one of the keyregulators of enzymes involved in metabolism and elimination of smallmolecules from the body. Activation of PXR is known to up-regulate orinduce the production of metabolic enzymes such as cytochrome P450 3A4(CYP3A4) as well as enzymes involved in transport such as OATP2 in theliver and intestine (Endocrine Reviews (2002) 23(5):687-702). When onedrug causes the up-regulation of these and other enzymes by activationof PXR, this can reduce the absorption and/or exposure of aco-administered drug that is susceptible to the up-regulated enzymes. Tominimize the risk of this type of drug-drug interaction, it is desirableto minimize PXR activation. Further, it is known that PXR is activatedby many different classes of molecules (Endocrine Reviews (2002)23(5):687-702). Thus for drugs that will be co-administered with otherdrugs, it is important to test for and minimize PXR activation.

Transporters have been identified as playing a role in thepharmacokinetic, safety and efficacy profile or drugs, and certaindrug-drug interactions are mediated by transporters. See, Giacomini K M,et al. ““Membrane transporters in drug development,” Nat. Rev DrugDiscov. 9: 215-236, 2010; Zhang L, et al. “Transporter-MediatedDrug-Drug Interactions,” Clin. Pharm. Ther. 89(4):481-484 (2011). Onetranporter, the organic cation transporter 2 (OCT2; SLC22A2), is amember of the solute carrier (SLC) super-family of transporters and isprimarily localized on the basolateral membrane of the renal proximaltubule. OCT2, in concert with apical expressed multidrug and toxinextrusion (MATE) transporters 1 and 2-K, is believed to form the majorcationic secretion pathway in the kidney and has been shown to transportendogenous compounds including creatinine and xenobiotics includingmetformin. Inhibition of OCT2 can thus lead to increased levels of serumcreatinine and the potential for increased levels of other OCT2substrates. It is important as well to test and reduce OCT2 inhibitionof drugs.

A goal of antiretroviral therapy is to achieve viral suppression in theHIV infected patient. Treatment guidelines published by the UnitedStates Department of Health and Human Services provide that achievementof viral suppression requires the use of combination therapies, i.e.,several drugs from at least two or more drug classes. (Panel onAntiretroviral Guidelines for Adults and Adolescents. Guidelines for theuse of antiretroviral agents in HIV-1-infected adults and adolescents.Department of Health and Human Services. Available athttp://aidsinfo.nih.gov/ContentFiles/AdultandAdolescentGL.pdf. Sectionaccessed Mar. 14, 2013.) In addition, decisions regarding the treatmentof HIV infected patients are complicated when the patient requirestreatment for other medical conditions (Id. at E-12). Because thestandard of care requires the use of multiple different drugs tosuppress HIV, as well as to treat other conditions the patient may beexperiencing, the potential for drug interaction is a criterion forselection of a drug regimen. As such, there is a need for antiretroviraltherapies having a decreased potential for drug interactions.

Accordingly, there is a need for new agents that inhibit the replicationof HIV and that minimize PXR activation when co-administered with otherdrugs.

BRIEF SUMMARY

The present invention is directed to novel polycyclic carbamoylpyridonecompounds, having antiviral activity, including stereoisomers andpharmaceutically acceptable salts thereof, and the use of such compoundsin the treatment of HIV infections. The compounds of the invention maybe used to inhibit the activity of HIV integrase and may be used toreduce HIV replication.

In one embodiment of the present invention, compounds having thefollowing Formula (I) are provided:

or a stereoisomer or pharmaceutically acceptable salt thereof,wherein:

X is —O— or —NZ³— or —CHZ³—;

W is —CHZ²—;

Z¹, Z² and Z³ are each, independently, hydrogen or C₁₋₃alkyl, or whereinZ¹ and Z² or Z¹ and Z³, taken together, form -L- wherein L is—C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂C(R^(a))₂—, or—C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—, wherein at least one of Z¹ andZ² or Z¹ and Z³, taken together, form -L-;

Z⁴ is a bond, —CH₂—, or —CH₂CH₂—;

Y¹ and Y² are each, independently, hydrogen, C₁₋₃alkyl or C₁₋₃haloalkyl;

R¹ is phenyl substituted with one to three halogens; and

each R^(a) is, independently, hydrogen, halo, hydroxyl or C₁₋₄alkyl.

In another embodiment of the present invention, compounds having thefollowing Formula (I) are provided:

or a stereoisomer or pharmaceutically acceptable salt thereof,wherein:

X is —O— or —NZ³— or —CHZ³—;

W is —O— or —NZ²— or —CHZ²—;

Z¹, Z² and Z³ are each, independently, hydrogen or C₁₋₃alkyl, or whereinZ¹ and Z² or Z¹ and Z³, taken together, form -L- wherein L is—C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂OC(R^(a))₂—,—C(R^(a))₂NR^(a)C(R^(a))₂—, —C(R^(a))₂SC(R^(a))₂—,—C(R^(a))₂S(O)C(R^(a))₂—, —C(R^(a))₂SO₂C(R^(a))₂—,—C(R^(a))₂OC(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂OC(R^(a))₂—,—C(R^(a))₂NR^(a)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂NR^(a)C(R^(a))₂—(R^(a)),—C(R^(a))₂SC(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂SC(R^(a))₂—,—C(R^(a))₂S(O)C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂S(O)C(R^(a))₂—,—C(R^(a))₂SO₂C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂SO₂C(R^(a))₂—,—C(R^(a))₂SO₂NR^(a)C(R^(a))₂— or —C(R^(a))₂NR^(a)SO₂C(R^(a))₂—;

Z⁴ is a bond or —CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—, —CH₂OCH₂—, —CH₂NR^(a)CH₂—,—CH₂SCH₂—, —CH₂S(O)CH₂— or —CH₂SO₂CH₂—;

Y¹ and Y² are each, independently, hydrogen, C₁₋₃alkyl or C₁₋₃haloalkyl,or Y¹ and Y², together with the carbon atom to which they are attached,form a carbocyclic ring having from 3 to 6 ring atoms or a heterocyclicring having from 3 to 6 ring atoms, wherein the carbocyclic orheterocyclic ring is optionally substituted with one or more R^(a);

R¹ is optionally substituted aryl or optionally substituted heteroaryl;and

each R^(a) is, independently, hydrogen, halo, hydroxyl or C₁₋₄alkyl, orwherein two R^(a) groups, together with the carbon atom to which theyare attached, form ═O, and

wherein at least one of: (i) Z¹ and Z² or Z¹ and Z³, taken together,form -L-; or (ii) Y¹ and Y², together with the carbon atom to which theyare attached, form a carbocyclic ring having from 3 to 6 ring atoms or aheterocyclic ring having from 3 to 6 ring atoms.

In another embodiment, a pharmaceutical composition is providedcomprising a compound having Formula (I), or a stereoisomer orpharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier, diluent or excipient.

The invention also provides the use of a pharmaceutical composition asdescribed hereinabove for the treatment of an HIV infection in a humanbeing having or at risk of having the infection.

In another embodiment, a method of using a compound having Formula (I)in therapy is provided. In particular, a method of treating theproliferation of the HIV virus, treating AIDS, or delaying the onset ofAIDS or ARC symptoms in a mammal (e.g., a human) is provided, comprisingadministering to the mammal a compound having Formula (I), or astereoisomer or pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier, diluent or excipient.

In another embodiment, use of a compound of Formula (I) as describedherein, or a pharmaceutically acceptable salt thereof, for the treatmentof an HIV infection in a human being having or at risk of having theinfection is disclosed.

In another embodiment, the use of a compound of Formula (I) as describedherein, or a pharmaceutically acceptable salt thereof, for themanufacture of a medicament for the treatment of an HIV infection in ahuman being having or at risk of having the infection is disclosed.

In another embodiment, an article of manufacture comprising acomposition effective to treat an HIV infection; and packaging materialcomprising a label which indicates that the composition can be used totreat infection by HIV is disclosed. Exemplary compositions comprise acompound of Formula (I) according to this invention or apharmaceutically acceptable salt thereof.

In still another embodiment, a method of inhibiting the replication ofHIV is disclosed. The method comprises exposing the virus to aneffective amount of the compound of Formula (I), or a salt thereof,under conditions where replication of HIV is inhibited.

In another embodiment, the use of a compound of Formula (I) to inhibitthe activity of the HIV integrase enzyme is disclosed.

In another embodiment, the use of a compound of Formula (I), or a saltthereof, to inhibit the replication of HIV is disclosed.

Other embodiments, objects, features and advantages will be set forth inthe detailed description of the embodiments that follows, and in partwill be apparent from the description, or may be learned by practice, ofthe claimed invention. These objects and advantages will be realized andattained by the processes and compositions particularly pointed out inthe written description and claims hereof. The foregoing Summary hasbeen made with the understanding that it is to be considered as a briefand general synopsis of some of the embodiments disclosed herein, isprovided solely for the benefit and convenience of the reader, and isnot intended to limit in any manner the scope, or range of equivalents,to which the appended claims are lawfully entitled.

DETAILED DESCRIPTION

In the following description, certain specific details are set forth inorder to provide a thorough understanding of various embodiments of theinvention. However, one skilled in the art will understand that theinvention may be practiced without these details. The description belowof several embodiments is made with the understanding that the presentdisclosure is to be considered as an exemplification of the claimedsubject matter, and is not intended to limit the appended claims to thespecific embodiments illustrated. The headings used throughout thisdisclosure are provided for convenience only and are not to be construedto limit the claims in any way. Embodiments illustrated under anyheading may be combined with embodiments illustrated under any otherheading.

Definitions

Unless the context requires otherwise, throughout the presentspecification and claims, the word “comprise” and variations thereof,such as, “comprises” and “comprising” are to be construed in an open,inclusive sense, that is as “including, but not limited to”.

Reference throughout this specification to “one embodiment” or “anembodiment” means that a particular feature, structure or characteristicdescribed in connection with the embodiment is included in at least oneembodiment of the present invention. Thus, the appearances of thephrases “in one embodiment” or “in an embodiment” in various placesthroughout this specification are not necessarily all referring to thesame embodiment. Furthermore, the particular features, structures, orcharacteristics may be combined in any suitable manner in one or moreembodiments.

Unless the context requires otherwise, reference throughout thisspecification to “a compound of Formula (I)” or “compounds of Formula(I)” refers to all embodiments of Formula (I), including, for example,compounds of Formulas (II-A), (II-B), (II-C), (III-A), (III-B), (III-C),(III-D), (III-E), (III-F), (III-G), (III-H), (IV-AA), (IV-AB), (IV-AC),(IV-AD), (IV-AE), (IV-AF), (IV-AG), (IV-AH), (TV-BA), (IV-BB), (IV-BC),(IV-BD), (IV-BE), (IV-BF), (IV-BG), and (IV-BH), as well as the specificcompounds disclosed herein.

“Amino” refers to the —NH₂ radical.

“Cyano” refers to the —CN radical.

“Hydroxy” or “hydroxyl” refers to the —OH radical.

“Imino” refers to the ═NH substituent.

“Nitro” refers to the —NO₂ radical.

“Oxo” refers to the ═O substituent.

“Thioxo” refers to the ═S substituent.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, which is saturated orunsaturated (i.e., contains one or more double and/or triple bonds),having from one to twelve carbon atoms (C₁-C₁₂ alkyl), preferably one toeight carbon atoms (C₁-C₈ alkyl) or one to six carbon atoms (C₁-C₆alkyl), and which is attached to the rest of the molecule by a singlebond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl),n-butyl, n-pentyl, 1,1-dimethylethyl (1-butyl), 3-methylhexyl,2-methylhexyl, ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl,penta-1,4-dienyl, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and thelike. Unless stated otherwise specifically in the specification, analkyl group may be optionally substituted.

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon chain linking the rest of the molecule to a radical group,consisting solely of carbon and hydrogen, which is saturated orunsaturated (i.e., contains one or more double and/or triple bonds), andhaving from one to twelve carbon atoms, e.g., methylene, ethylene,propylene, n-butylene, ethenylene, propenylene, n-butenylene,propynylene, n-butynylene, and the like. The alkylene chain is attachedto the rest of the molecule through a single or double bond and to theradical group through a single or double bond. The points of attachmentof the alkylene chain to the rest of the molecule and to the radicalgroup can be through one carbon or any two carbons within the chain.Unless stated otherwise specifically in the specification, an alkylenechain may be optionally substituted.

“Alkoxy” refers to a radical of the formula —OR_(A) where R_(A) is analkyl radical as defined above containing one to twelve carbon atoms.Unless stated otherwise specifically in the specification, an alkoxygroup may be optionally substituted.

“Alkylamino” refers to a radical of the formula —NHR_(A) or —NR_(A)R_(A)where each R_(A) is, independently, an alkyl radical as defined abovecontaining one to twelve carbon atoms. Unless stated otherwisespecifically in the specification, an alkylamino group may be optionallysubstituted.

Thioalkyl” refers to a radical of the formula —SR_(A) where R_(A) is analkyl radical as defined above containing one to twelve carbon atoms.Unless stated otherwise specifically in the specification, a thioalkylgroup may be optionally substituted.

“Aryl” refers to a monocylic hydrocarbon ring system radical comprisinghydrogen and 6 to 18 carbon atoms. Aryl radicals include, but are notlimited to, aryl radicals derived from benzene. Unless stated otherwisespecifically in the specification, the term “aryl” or the prefix “ar-”(such as in “aralkyl”) is meant to include aryl radicals that areoptionally substituted.

“Aralkyl” refers to a radical of the formula —R_(B)—R_(C) where R_(B) isan alkylene chain as defined above and R_(C) is one or more arylradicals as defined above, for example, benzyl. Unless stated otherwisespecifically in the specification, an aralkyl group may be optionallysubstituted.

“Cycloalkyl” or “carbocyclic ring” refers to a stable non-aromaticmonocyclic hydrocarbon radical consisting solely of carbon and hydrogenatoms, having from three to fifteen carbon atoms, preferably having fromthree to ten carbon atoms, and which is saturated or unsaturated andattached to the rest of the molecule by a single bond. Monocyclicradicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl. Unless otherwise statedspecifically in the specification, a cycloalkyl group may be optionallysubstituted.

“Cycloalkylalkyl” refers to a radical of the formula —R_(B)R_(D) whereR_(B) is an alkylene chain as defined above and R_(D) is a cycloalkylradical as defined above. Unless stated otherwise specifically in thespecification, a cycloalkylalkyl group may be optionally substituted.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodo.

“Haloalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more halo radicals, as defined above, e.g.,trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl,1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and thelike. Unless stated otherwise specifically in the specification, ahaloalkyl group may be optionally substituted.

“Heterocyclyl” or “heterocyclic ring” refers to a stable 3- to18-membered non-aromatic ring radical which consists of two to twelvecarbon atoms and from one to six heteroatoms selected from the groupconsisting of nitrogen, oxygen and sulfur. In the embodiments disclosedherein, the heterocyclyl radical is a monocyclic ring system; and theheterocyclyl radical may be partially or fully saturated. Examples ofsuch heterocyclyl radicals include, but are not limited to, dioxolanyl,thienyl, [1,3]dithianyl, imidazolinyl, imidazolidinyl, isothiazolidinyl,isoxazolidinyl, morpholinyl, 2-oxopiperazinyl, 2-oxopiperidinyl,2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl,4-piperidonyl, pyrrolidinyl, pyrazolidinyl, thiazolidinyl,tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl,thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.Unless stated otherwise specifically in the specification, aheterocyclyl group may be optionally substituted.

“N-heterocyclyl” refers to a heterocyclyl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heterocyclyl radical to the rest of the molecule is through anitrogen atom in the heterocyclyl radical. Unless stated otherwisespecifically in the specification, an N-heterocyclyl group may beoptionally substituted.

“Heterocyclylalkyl” refers to a radical of the formula —R_(B)R_(E) whereR_(B) is an alkylene chain as defined above and R_(E) is a heterocyclylradical as defined above, and if the heterocyclyl is anitrogen-containing heterocyclyl, the heterocyclyl may be attached tothe alkyl radical at the nitrogen atom. Unless stated otherwisespecifically in the specification, a heterocyclylalkyl group may beoptionally substituted.

“Heteroaryl” refers to a 5- to 14-membered monocyclic ring systemradical comprising hydrogen atoms, one to thirteen carbon atoms, one tosix heteroatoms selected from the group consisting of nitrogen, oxygenand sulfur. Examples include, but are not limited to, azepinyl, furanyl,furanonyl, isothiazolyl, imidazolyl, isoxazolyl, oxadiazolyl,2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl,1-oxidopyrazinyl, 1-oxidopyridazinyl, pyrazolyl, pyridinyl, pyrazinyl,pyrimidinyl, pyridazinyl, thiazolyl, thiadiazolyl, triazolyl,tetrazolyl, triazinyl, thiophenyl, and thienyl. Unless stated otherwisespecifically in the specification, a heteroaryl group may be optionallysubstituted.

“N-heteroaryl” refers to a heteroaryl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heteroaryl radical to the rest of the molecule is through a nitrogenatom in the heteroaryl radical. Unless stated otherwise specifically inthe specification, an N-heteroaryl group may be optionally substituted.

“Heteroarylalkyl” refers to a radical of the formula —R_(B)R_(F) whereR_(B) is an alkylene chain as defined above and R_(F) is a heteroarylradical as defined above. Unless stated otherwise specifically in thespecification, a heteroarylalkyl group may be optionally substituted.The term “substituted” used herein means any of the above groups (i.e.,alkyl, alkylene, alkoxy, alkylamino, thioalkyl, aryl, aralkyl,cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl,heterocyclylalkyl, heteroaryl, N-heteroaryl and/or heteroarylalkyl)wherein at least one hydrogen atom is replaced by a bond to anon-hydrogen atoms such as, but not limited to: a halogen atom such asF, Cl, Br, and I; an oxygen atom in groups such as hydroxyl groups,alkoxy groups, and ester groups; a sulfur atom in groups such as thiolgroups, thioalkyl groups, sulfone groups, sulfonyl groups, and sulfoxidegroups; a nitrogen atom in groups such as amines, amides, alkylamines,dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides,imides, and enamines; a silicon atom in groups such as trialkylsilylgroups, dialkylarylsilyl groups, alkyldiarylsilyl groups, andtriarylsilyl groups; and other heteroatoms in various other groups.“Substituted” also means any of the above groups in which one or morehydrogen atoms are replaced by a higher-order bond (e.g., a double- ortriple-bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl,and ester groups; and nitrogen in groups such as imines, oximes,hydrazones, and nitriles. For example, “substituted” includes any of theabove groups in which one or more hydrogen atoms are replaced with—NR_(G)R_(H), —NR_(G)C(═O)R_(H), —NR_(G)C(═O)NR_(G)R_(H),—NR_(G)C(═O)OR_(H), —NR_(G)C(═NR_(g))NR_(G)R_(H), —NR_(G)SO₂R_(H),—OC(═O)NR_(G)R_(H), —OR_(G), —SR_(G). —SOR_(G), —SO₂R_(G), —OSO₂R_(G),—SO₂OR_(G), ═NSO₂R_(G), and —SO₂NR_(G)R_(H). “Substituted also means anyof the above groups in which one or more hydrogen atoms are replacedwith —C(═O)R_(G), —C(═O)OR_(G), —C(═O)NR_(G)R_(H), —CH₂SO₂R_(G),—CH₂SO₂NR_(G)R_(H). In the foregoing, R_(G) and R_(H) are the same ordifferent and independently hydrogen, alkyl, alkoxy, alkylamino,thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl,heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl,N-heteroaryl and/or heteroarylalkyl. “Substituted” further means any ofthe above groups in which one or more hydrogen atoms are replaced by abond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo,alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl,heterocyclylalkyl, heteroaryl, N-heteroaryl and/or heteroarylalkylgroup. In addition, each of the foregoing substituents may also beoptionally substituted with one or more of the above substituents.

The term “protecting group,” as used herein, refers to a labile chemicalmoiety which is known in the art to protect reactive groups includingwithout limitation, hydroxyl and amino groups, against undesiredreactions during synthetic procedures. Hydroxyl and amino groupsprotected with a protecting group are referred to herein as “protectedhydroxyl groups” and “protected amino groups”, respectively. Protectinggroups are typically used selectively and/or orthogonally to protectsites during reactions at other reactive sites and can then be removedto leave the unprotected group as is or available for further reactions.Protecting groups as known in the art are described generally in Greeneand Wuts, Protective Groups in Organic Synthesis, 3rd edition, JohnWiley & Sons, New York (1999). Generally, groups are protected orpresent as a precursor that will be inert to reactions that modify otherareas of the parent molecule for conversion into their final groups atan appropriate time. Further representative protecting or precursorgroups are discussed in Agrawal, et al., Protocols for OligonucleotideConjugates, Eds, Humana Press; New Jersey, 1994; Vol. 26 pp. 1-72.Examples of “hydroxyl protecting groups” include, but are not limitedto, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-trimethylsilylethyl,p-chlorophenyl, 2,4-dinitrophenyl, benzyl, 2,6-dichlorobenzyl,diphenylmethyl, p-nitrobenzyl, triphenylmethyl, trimethylsilyl,triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl (TBDPS),triphenylsilyl, benzoylformate, acetate, chloroacetate,trichloroacetate, trifluoroacetate, pivaloate, benzoate,p-phenylbenzoate, 9-fluorenylmethyl carbonate, mesylate and tosylate.Examples of “amino protecting groups” include, but are not limited to,carbamate-protecting groups, such as 2-trimethylsilylethoxycarbonyl(Teoc), 1-methyl-1-(4-biphenylyl)ethoxycarbonyl (Bpoc), t-butoxycarbonyl(BOC), allyloxycarbonyl (Alloc), 9-fluorenylmethyloxycarbonyl (Fmoc),and benzyloxycarbonyl (Cbz); amide protecting groups, such as formyl,acetyl, trihaloacetyl, benzoyl, and nitrophenylacetyl;sulfonamide-protecting groups, such as 2-nitrobenzenesulfonyl; and imineand cyclic imide protecting groups, such as phthalimido anddithiasuccinoyl.

The invention disclosed herein is also meant to encompass allpharmaceutically acceptable compounds of Formula (I) beingisotopically-labeled by having one or more atoms replaced by an atomhaving a different atomic mass or mass number. Examples of isotopes thatcan be incorporated into the disclosed compounds include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, andiodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³¹P,³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I, respectively. These radiolabeledcompounds could be useful to help determine or measure the effectivenessof the compounds, by characterizing, for example, the site or mode ofaction, or binding affinity to pharmacologically important site ofaction. Certain isotopically-labeled compounds of Formula (I), forexample, those incorporating a radioactive isotope, are useful in drugand/or substrate tissue distribution studies. The radioactive isotopestritium, i.e. ³H, and carbon-14, i.e. ¹⁴C, are particularly useful forthis purpose in view of their ease of incorporation and ready means ofdetection.

Substitution with heavier isotopes such as deuterium, i.e. ²H, mayafford certain therapeutic advantages resulting from greater metabolicstability. For example, in vivo half-life may increase or dosagerequirements may be reduced. Thus, heavier isotopes may be preferred insome circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹³O and¹³N, can be useful in Positron Emission Topography (PET) studies forexamining substrate receptor occupancy. Isotopically-labeled compoundsof Formula (I) can generally be prepared by conventional techniquesknown to those skilled in the art or by processes analogous to thosedescribed in the Examples as set out below using an appropriateisotopically-labeled reagent in place of the non-labeled reagentpreviously employed.

The invention disclosed herein is also meant to encompass the in vivometabolic products of the disclosed compounds. Such products may resultfrom, for example, the oxidation, reduction, hydrolysis, amidation,esterification, and the like of the administered compound, primarily dueto enzymatic processes. Accordingly, the invention includes compoundsproduced by a process comprising administering a compound of thisinvention to a mammal for a period of time sufficient to yield ametabolic product thereof. Such products are typically identified byadministering a radiolabeled compound of the invention in a detectabledose to an animal, such as rat, mouse, guinea pig, monkey, or to human,allowing sufficient time for metabolism to occur, and isolating itsconversion products from the urine, blood or other biological samples.

“Stable compound” and “stable structure” are meant to indicate acompound that is sufficiently robust to survive isolation to a usefuldegree of purity from a reaction mixture, and formulation into anefficacious therapeutic agent.

“Mammal” includes humans and both domestic animals such as laboratoryanimals and household pets (e.g., cats, dogs, swine, cattle, sheep,goats, horses, rabbits), and non-domestic animals such as wildlife andthe like.

“Optional” or “optionally” means that the subsequently described eventof circumstances may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not. For example, “optionally substituted aryl” means that thearyl radical may or may not be substituted and that the descriptionincludes both substituted aryl radicals and aryl radicals having nosubstitution.

“Pharmaceutically acceptable carrier, diluent or excipient” includeswithout limitation any adjuvant, carrier, excipient, glidant, sweeteningagent, diluent, preservative, dye/colorant, flavor enhancer, surfactant,wetting agent, dispersing agent, suspending agent, stabilizer, isotonicagent, solvent, or emulsifier which has been approved by the UnitedStates Food and Drug Administration as being acceptable for use inhumans or domestic animals.

“Pharmaceutically acceptable salt” refers to a salt of a compound thatis pharmaceutically acceptable and that possesses (or can be convertedto a form that possesses) the desired pharmacological activity of theparent compound. Examples of “pharmaceutically acceptable salts” of thecompounds disclosed herein include salts derived from an appropriatebase, such as an alkali metal (for example, sodium), an alkaline earthmetal (for example, magnesium), ammonium and NX₄ ⁺ (wherein X is C₁-C₄alkyl). Pharmaceutically acceptable salts of a nitrogen atom or an aminogroup include for example salts of organic carboxylic acids such asacetic, benzoic, camphorsulfonic, citric, glucoheptonic, gluconic,lactic, fumaric, tartaric, maleic, malonic, malic, mandelic, isethionic,lactobionic, succinic, 2-napththalenesulfonic, oleic, palmitic,propionic, stearic, and trimethylacetic acids; organic sulfonic acids,such as methanesulfonic, ethanesulfonic, benzenesulfonic andp-toluenesulfonic acids; and inorganic acids, such as hydrochloric,hydrobromic, sulfuric, nitric, phosphoric and sulfamic acids.Pharmaceutically acceptable salts of a compound of a hydroxy groupinclude the anion of said compound in combination with a suitable cationsuch as Na⁺ and NX₄ ⁺ (wherein X is independently selected from H or aC₁-C₄ alkyl group). Pharmaceutically acceptable salts also include saltsformed when an acidic proton present in the parent compound is replacedby either a metal ion, e.g., an alkali metal ion, an alkaline earth ion,or an aluminum ion; or coordinates with an organic base such asdiethanolamine, triethanolamine, N-methylglucamine and the like. Alsoincluded in this definition are ammonium and substituted or quaternizedammonium salts. Representative non-limiting lists of pharmaceuticallyacceptable salts can be found in S. M. Berge et al., J. Pharma Sci.,66(1), 1-19 (1977), and Remington: The Science and Practice of Pharmacy,R. Hendrickson, ed., 21st edition, Lippincott, Williams & Wilkins,Philadelphia, Pa., (2005), at p. 732, Table 38-5, both of which arehereby incorporated by reference herein.

For therapeutic use, salts of active ingredients of the compoundsdisclosed herein will typically be pharmaceutically acceptable, i.e.they will be salts derived from a physiologically acceptable acid orbase. However, salts of acids or bases which are not pharmaceuticallyacceptable may also find use, for example, in the preparation orpurification of a compound of Formula (I) or another compound of theinvention. All salts, whether or not derived from a physiologicallyacceptable acid or base, are within the scope of the present invention.

Metal salts typically are prepared by reacting the metal hydroxide witha compound of this invention. Examples of metal salts which are preparedin this way are salts containing Li⁺, Na⁺, and K⁺. A less soluble metalsalt can be precipitated from the solution of a more soluble salt byaddition of the suitable metal compound.

In addition, salts may be formed from acid addition of certain organicand inorganic acids, e.g., HCl, HBr, H₂SO₄, H₃PO₄ or organic sulfonicacids, to basic centers, typically amines. Finally, it is to beunderstood that the compositions herein comprise compounds disclosedherein in their un-ionized, as well as zwitterionic form, andcombinations with stoichiometric amounts of water as in hydrates.

Often crystallizations produce a solvate of the compound of theinvention. As used herein, the term “solvate” refers to an aggregatethat comprises one or more molecules of a compound of the invention withone or more molecules of solvent. The solvent may be water, in whichcase the solvate may be a hydrate. Alternatively, the solvent may be anorganic solvent. Thus, the compounds of the present invention may existas a hydrate, including a monohydrate, dihydrate, hemihydrate,sesquihydrate, trihydrate, tetrahydrate and the like, as well as thecorresponding solvated forms. The compound of the invention may be truesolvates, while in other cases, the compound of the invention may merelyretain adventitious water or be a mixture of water plus someadventitious solvent.

A “pharmaceutical composition” refers to a formulation of a compound ofthe invention and a medium generally accepted in the art for thedelivery of the biologically active compound to mammals, e.g., humans.Such a medium includes all pharmaceutically acceptable carriers,diluents or excipients therefor.

“Effective amount” or “therapeutically effective amount” refers to anamount of a compound according to the invention, which when administeredto a patient in need thereof, is sufficient to effect treatment fordisease-states, conditions, or disorders for which the compounds haveutility. Such an amount would be sufficient to elicit the biological ormedical response of a tissue system, or patient that is sought by aresearcher or clinician. The amount of a compound according to theinvention which constitutes a therapeutically effective amount will varydepending on such factors as the compound and its biological activity,the composition used for administration, the time of administration, theroute of administration, the rate of excretion of the compound, theduration of the treatment, the type of disease-state or disorder beingtreated and its severity, drugs used in combination with orcoincidentally with the compounds of the invention, and the age, bodyweight, general health, sex and diet of the patient. Such atherapeutically effective amount can be determined routinely by one ofordinary skill in the art having regard to their own knowledge, thestate of the art, and this disclosure.

The term “treatment” as used herein is intended to mean theadministration of a compound or composition according to the presentinvention to alleviate or eliminate symptoms of HIV infection and/or toreduce viral load in a patient. The term “treatment” also encompassesthe administration of a compound or composition according to the presentinvention post-exposure of the individual to the virus but before theappearance of symptoms of the disease, and/or prior to the detection ofthe virus in the blood, to prevent the appearance of symptoms of thedisease and/or to prevent the virus from reaching detectible levels inthe blood, and the administration of a compound or composition accordingto the present invention to prevent perinatal transmission of HIV frommother to baby, by administration to the mother before giving birth andto the child within the first days of life.

The term “antiviral agent” as used herein is intended to mean an agent(compound or biological) that is effective to inhibit the formationand/or replication of a virus in a human being, including but notlimited to agents that interfere with either host or viral mechanismsnecessary for the formation and/or replication of a virus in a humanbeing.

The term “inhibitor of HIV replication” as used herein is intended tomean an agent capable of reducing or eliminating the ability of HIV toreplicate in a host cell, whether in vitro, ex vivo or in vivo.

The compounds of the invention, or their pharmaceutically acceptablesalts may contain one or more asymmetric centers and may thus give riseto enantiomers, diastereomers, and other stereoisomeric forms that maybe defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as(D)- or (L)- for amino acids. The present invention is meant to includeall such possible isomers, as well as their racemic and optically pureforms. Optically active (+) and (−), (R)- and (S)-, or (D)- and(L)-isomers may be prepared using chiral synthons or chiral reagents, orresolved using conventional techniques, for example, chromatography andfractional crystallization. Conventional techniques for thepreparation/isolation of individual enantiomers include chiral synthesisfrom a suitable optically pure precursor or resolution of the racemate(or the racemate of a salt or derivative) using, for example, chiralhigh pressure liquid chromatography (HPLC). When the compounds describedherein contain olefinic double bonds or other centres of geometricasymmetry, and unless specified otherwise, it is intended that thecompounds include both E and Z geometric isomers. Likewise, alltautomeric forms are also intended to be included.

A “stereoisomer” refers to a compound made up of the same atoms bondedby the same bonds but having different three-dimensional structures,which are not interchangeable. The present invention contemplatesvarious stereoisomers and mixtures thereof and includes “enantiomers”,which refers to two stereoisomers whose molecules are nonsuperimposeablemirror images of one another.

A “tautomer” refers to a proton shift from one atom of a molecule toanother atom of the same molecule. The present invention includestautomers of any said compounds.

A “prodrug” refers to a compound that is chemically designed toefficiently liberate the parent drug after overcoming biologicalbarriers to oral delivery. In certain embodiments, the present inventionincludes prodrugs of the compounds of Formula (I).

Compounds

As noted above, in one embodiment of the present invention, compoundshaving antiviral activity are provided, the compounds having thefollowing Formula (I):

or a stereoisomer or pharmaceutically acceptable salt thereof,

wherein:

X is —O— or —NZ³— or —CHZ³—;

W is —CHZ²—;

Z¹, Z² and Z³ are each, independently, hydrogen or C₁₋₃alkyl, or whereinZ¹ and Z² or Z¹ and Z³, taken together, form -L- wherein L is—C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂C(R^(a))₂—, or—C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—, wherein at least one of Z¹ andZ² or Z¹ and Z³, taken together, form -L-;

Z⁴ is a bond, —CH₂—, or —CH₂CH₂—;

Y¹ and Y² are each, independently, hydrogen, C₁₋₃alkyl or C₁₋₃haloalkyl;

R¹ is phenyl substituted with one to three halogens; and

each R^(a) is, independently, hydrogen, halo, hydroxyl or C₁₋₄alkyl.

In another embodiment, compounds are provided having the followingFormula (II-A):

In another embodiment, compounds are provided having the followingFormula (II-B):

In another embodiment, compounds are provided having the followingFormula (II-C):

In another embodiment, L is —C(R^(a))₂—. In a further embodiment, L is—C(R^(a))₂C(R^(a))₂—. In still a further embodiment, L is—C(R^(a))₂C(R^(a))₂C(R^(a))₂—. In still a further embodiment, each R^(a)is hydrogen. In still a further embodiment, one R^(a) is methyl and eachremaining R^(a) is hydrogen. In still a further embodiment, one R^(a) ishalogen and each remaining R^(a) is hydrogen. In still a furtherembodiment, two R^(a) are halogen and each remaining R^(a) is hydrogen.In still a further embodiment, one R^(a) is halogen and each remainingR^(a) is hydrogen.

In another embodiment, X is —O—. In another embodiment, X is —NZ³—. Inanother embodiment, X is —NH—. 16. In another embodiment, X is —CHZ³—and Z¹ and Z³, taken together, form -L-. In a further embodiment, Z² ishydrogen. In another embodiment, X is —CH₂—.

In another embodiment, Z⁴ is a bond or —CH₂—. In another embodiment, Z⁴is —CH₂—. In another embodiment, Z⁴ is a bond.

In another embodiment, Y¹ and Y² are each independently hydrogen, methylor trifluoromethyl.

In another embodiment, R¹ is substituted with one halogen. In a furtherembodiment, R¹ is 4-fluorophenyl or 2-fluorophenyl.

In another embodiment, R¹ is substituted with two halogens. In a furtherembodiment, R¹ is 2,4-difluorophenyl, 2,3-difluorophenyl,2,6-difluorophenyl, 3-fluoro-4-chlorophenyl, 3,4-difluorophenyl,2-fluoro-4-chlorophenyl, or 3,5-difluorophenyl. In still a furtherembodiment, R¹ is 2,4-difluorophenyl.

In another embodiment, R¹ is substituted with three halogens. In afurther embodiment, R¹ is 2,4,6-trifluorophenyl or2,3,4-trifluorophenyl. In still a further embodiment, R¹ is2,4,6-trifluorophenyl.

In one embodiment, a pharmaceutical composition is provided comprising acompound of any one of the Formulas (I), (II-A), (II-B), or (1-C), asnoted above, or a stereoisomer or pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier, diluent orexcipient.

Another embodiment is provided comprising a method of treating an HIVinfection in a human having or at risk of having the infection byadministering to the human a therapeutically effective amount of acompound of any one of the Formulas (I), (II-A), (II-B), or (II-C), asnoted above, or a pharmaceutical composition thereof. Another embodimentis provided comprising a method of treating or preventing an HIVinfection in a human having or at risk of having the infection byadministering to the human a therapeutically effective amount of acompound of any one of the Formulas (I), (II-A), (II-B), or (II-C), asnoted above, or a pharmaceutical composition thereof.

In another embodiment, the use of a compound of any one of the Formulas(I), (II-A), (II-B), or (II-C), as noted above, or a pharmaceuticalcomposition thereof, for the treatment of an HIV infection in a humanhaving or at risk of having the infection is provided. In anotherembodiment, the use of a compound of any one of the Formulas (I),(II-A), (II-B), or (II-C), as noted above, or a pharmaceuticalcomposition thereof, for the treatment or prevention of an HIV infectionin a human having or at risk of having the infection is provided.

In another embodiment, the use in medical therapy of a compound of anyone of the Formulas (I), (II-A), (II-B), or (II-C), as noted above, or apharmaceutical composition thereof, is provided.

In another embodiment, the use of a compound of any one of the Formulas(I), (II-A), (II-B), or (II-C), as noted above, or a pharmaceuticalcomposition thereof, for use in the therapeutic treatment of an HIVinfection is provided. In another embodiment, the use of a compound ofany one of the Formulas (I), (II-A), (II-B), or (II-C), as noted above,or a pharmaceutical composition thereof, for use in the prophylactic ortherapeutic treatment of an HIV infection is provided.

As further noted above, in another embodiment of the present invention,compounds having antiviral activity are provided, the compounds havingthe following Formula (I):

or a stereoisomer or pharmaceutically acceptable salt thereof,

wherein:

X is —O— or —NZ³— or —CHZ³—;

W is —O— or —NZ²— or —CHZ²—;

Z¹, Z² and Z³ are each, independently, hydrogen, C₁₋₃alkyl orC₁₋₃haloalkyl, or wherein Z¹ and Z² or Z¹ and Z³, taken together, form-L- wherein L is —C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂—,—C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂—,—C(R^(a))₂SO₂C(R^(a))₂—, —C(R^(a))₂OC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂NR^(a)C(R^(a))₂—, —C(R^(a))₂SC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂S(O)C(R^(a))₂—, —C(R^(a))₂SO₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂SO₂C(R^(a))₂—, —(SO₂C(R^(a))₂—, —C(R^(a))₂SO₂NR^(a)C(R^(a))₂—or —C(R^(a))₂NR^(a)SO₂C(R^(a))₂—;

Z⁴ is a bond or —CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—, —CH₂OCH₂—, —CH₂NR^(a)CH₂—,—CH₂SCH₂—, —CH₂S(O)CH₂— or —CH₂SO₂CH₂—;

Y¹ and Y² are each, independently, hydrogen or C₁₋₃alkyl, or Y¹ and Y²,together with the carbon atom to which they are attached, form acarbocyclic ring having from 3 to 6 ring atoms or a heterocyclic ringhaving from 3 to 6 ring atoms, wherein the carbocyclic or heterocyclicring is optionally substituted with one or more R^(a);

R¹ is optionally substituted aryl or optionally substituted heteroaryl;and

each R^(a) is, independently, hydrogen, halo, hydroxyl or C₁₋₄alkyl, orwherein two R^(a) groups, together with the carbon atom to which theyare attached, form ═O, and

wherein at least one of: (i) Z¹ and Z² or Z¹ and Z³, taken together,form -L-; or (ii) Y¹ and Y², together with the carbon atom to which theyare attached, form a carbocyclic ring having from 3 to 6 ring atoms or aheterocyclic ring having from 3 to 6 ring atoms.

In another embodiment, W is —CHZ²—.

In another embodiment, Z¹ and Z² or Z¹ and Z³, taken together, form -L-.

In another embodiment, compounds are provided having one of thefollowing Formulas (II-A), (II-B), or (II-C):

wherein L is —C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂—,—C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂—,—C(R^(a))₂SO₂C(R^(a))₂—, —C(R^(a))₂OC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂NR^(a)C(R^(a))₂—, —C(R^(a))₂SC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂S(O)C(R^(a))₂—, —C(R^(a))₂SO₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂SO₂C(R^(a))₂—, —C(R^(a))₂SO₂NR^(a)C(R^(a))₂— or—C(R^(a))₂NR^(a)SO₂C(R^(a))₂—.

In another embodiment, Y¹ and Y², together with the carbon atom to whichthey are attached, form a carbocyclic ring having from 3 to 6 ring atomsor a heterocyclic ring having from 3 to 6 ring atoms.

In another embodiment, compounds are provided having one of thefollowing Formulas (III-A), (III-B), (III-C) or (III-D):

wherein Z¹ and Z³ are each, independently, hydrogen or C₁₋₃alkyl.

In another embodiment, compounds are provided having one of thefollowing Formulas (III-E), (III-F), (III-G) or (III-H):

wherein Z and Z³ are each, independently, hydrogen or C₁₋₃alkyl.

In another embodiment, both (i) Z¹ and Z² or Z¹ and Z³, taken together,form -L-, and (ii) Y¹ and Y², together with the carbon atom to whichthey are attached, form a carbocyclic ring having from 3 to 6 ring atomsor a heterocyclic ring having from 3 to 6 ring atoms.

In another embodiment, compounds are provided having one of thefollowing Formulas (IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE), (IV-AF),(IV-AG) or (IV-AH):

wherein L is —C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂—,—C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂—,—C(R^(a))₂SO₂C(R^(a))₂—, —C(R^(a))₂OC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂NR^(a)C(R^(a))₂—, —C(R^(a))₂SC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂S(O)C(R^(a))₂—, —C(R^(a))₂SO₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂SO₂C(R^(a))₂—, —C(R^(a))₂SO₂NR^(a)C(R^(a))₂— or—C(R^(a))₂NR^(a)SO₂C(R^(a))₂—.

In another embodiment, compounds are provided having one of thefollowing Formulas (IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE), (IV-BF),(IV-BG) or (IV-BH):

wherein L is —C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂—,—C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂—,—C(R^(a))₂SO₂C(R^(a))₂—, —C(R^(a))₂OC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂OC(R^(a))₂—, —C(R^(a))₂NR^(a)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂NR^(a)C(R^(a))₂—, —C(R^(a))₂SC(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂SC(R^(a))₂—, —C(R^(a))₂S(O)C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂S(O)C(R^(a))₂—, —C(R^(a))₂SO₂C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂SO₂C(R^(a))₂—, —C(R^(a))₂SO₂NR^(a)C(R^(a))₂— or—C(R^(a))₂NR^(a)SO₂C(R^(a))₂—.

In another embodiment, L is —C(R^(a))₂—, —C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂C(R^(a))₂—, or—C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—. In a further embodiment, L is—C(R^(a))₂—. In still a further embodiment, L is —C(R^(a))₂C(R^(a))₂—.In still a further embodiment, L is —C(R^(a))₂C(R^(a))₂C(R^(a))₂—. Instill a further embodiment, each R^(a) is hydrogen. In still a furtherembodiment, one R^(a) is methyl and each remaining R^(a) is hydrogen. Instill a further embodiment, one R^(a) is halogen and each remainingR^(a) is hydrogen. In still a further embodiment, two R^(a) are halogenand each remaining R^(a) is hydrogen. In still a further embodiment, oneR^(a) is halogen and each remaining R^(a) is hydrogen.

In another embodiment, L is —C(R^(a))₂OC(R^(a))₂—,—C(R^(a))₂NR^(a)C(R^(a))₂—, —C(R^(a))₂SC(R^(a))₂—,—C(R^(a))₂S(O)C(R^(a))₂—, or —C(R^(a))₂SO₂C(R^(a))₂—. In a furtherembodiment, L is —C(R^(a))₂OC(R^(a))₂—. In still a further embodiment,each R^(a) is hydrogen. In still a further embodiment, one R^(a) ismethyl and each remaining R^(a) is hydrogen. In still a furtherembodiment, one R^(a) is halogen and each remaining R^(a) is hydrogen.In still a further embodiment, two R^(a) are halogen and each remainingR^(a) is hydrogen. In still a further embodiment, one R^(a) is halogenand each remaining R^(a) is hydrogen.

In another embodiment, X is —O—. In a further embodiment, Z² ishydrogen. In another embodiment, X is —NZ³—. In another embodiment, X is—NH—. In another embodiment, X is —CHZ³—. In another embodiment, X is—CH₂—.

In another embodiment, Z⁴ is a bond or —CH₂—. In another embodiment, Z⁴is —CH₂—. In another embodiment, Z⁴ is a bond.

In another embodiment, Y¹ and Y² are each independently hydrogen, methylor trifluoromethyl.

In another embodiment, R¹ is substituted with one halogen. In a furtherembodiment, R¹ is 4-fluorophenyl or 2-fluorophenyl.

In another embodiment, R¹ is phenyl. In another embodiment, R¹ ispyridinyl.

In another embodiment, R¹ is substituted with at least one halogen.

In another embodiment, R¹ is substituted with one halogen. In a furtherembodiment, R¹ is 4-fluorophenyl or 2-fluorophenyl.

In another embodiment, R¹ is substituted with two halogens. In a furtherembodiment, R¹ is 2,4-difluorophenyl, 2,3-difluorophenyl,2,6-difluorophenyl, 3-fluoro-4-chlorophenyl, 3,4-difluorophenyl,2-fluoro-4-chlorophenyl, or 3,5-difluorophenyl. In still a furtherembodiment, R¹ is 2,4-difluorophenyl.

In another embodiment, R¹ is substituted with three halogens. In afurther embodiment, R¹ is 2,4,6-trifluorophenyl or2,3,4-trifluorophenyl. In still a further embodiment, R¹ is2,4,6-trifluorophenyl.

In another embodiment, R¹ is 3-trifluoromethyl-4-fluorophenyl or2-cyclopropoxy-4-fluorophenyl.

In one embodiment, a pharmaceutical composition is provided comprising acompound of any one of Formulas (I), (II-A), (II-B), (II-C), (III-A),(III-B), (III-C), (III-D), (III-E), (III-F), (III-G), (III-H), (IV-AA),(IV-AB), (IV-AC), (IV-AD), (IV-AE), (IV-AF), (IV-AG), (IV-AH), (IV-BA),(IV-BB), (IV-BC), (IV-BD), (IV-BE), (IV-BF), (IV-BG), and (IV-BH), asnoted above, or a stereoisomer or pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier, diluent orexcipient.

Another embodiment is provided comprising a method of treating an HIVinfection in a human having or at risk of having the infection byadministering to the human a therapeutically effective amount of acompound of any one of Formulas (I), (II-A), (II-B), (II-C), (III-A),(III-B), (III-C), (III-D), (II-E), (III-F), (III-G), (III-H), (IV-AA),(IV-AB), (IV-AC), (IV-AD), (IV-AE), (IV-AF), (IV-AG), (IV-AH), (IV-BA),(IV-BB), (IV-BC), (IV-BD), (IV-BE), (IV-BF), (IV-BG), and (IV-BH), asnoted above, or a pharmaceutical composition thereof. Another embodimentis provided comprising a method of treating or preventing an HIVinfection in a human having or at risk of having the infection byadministering to the human a therapeutically effective amount of acompound of any one of Formulas (I), (II-A), (II-B), (II-C), (III-A),(III-B), (III-C), (I-D), (III-E), (III-F), (III-G), (II-H), (IV-AA),(IV-AB), (IV-AC), (IV-AD), (IV-AE), (IV-AF), (IV-AG), (IV-AH), (IV-BA),(IV-BB), (IV-BC), (IV-BD), (IV-BE), (IV-BF), (IV-BG), and (IV-BH), asnoted above, or a pharmaceutical composition thereof.

In another embodiment, the use of a compound of any one of Formulas (I),(II-A), (II-B), (II-C), (III-A), (III-B), (III-C), (III-D), (III-E),(III-F), (III-G), (III-H), (IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE),(IV-AF), (IV-AG), (IV-AH), (IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE),(IV-BF), (IV-BG), and (IV-BH), as noted above, or a pharmaceuticalcomposition thereof for the treatment of an HIV infection in a humanhaving or at risk of having the infection. In another embodiment, theuse of a compound of any one of Formulas (I), (II-A), (II-B), (II-C),(III-A), (III-B), (III-C), (III-D), (III-E), (III-F), (III-G), (III-H),(IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE), (IV-AF), (IV-AG), (IV-AH),(IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE), (IV-BF), (IV-BG), and(IV-BH), as noted above, or a pharmaceutical composition thereof for thetreatment or prevention of an HIV infection in a human having or at riskof having the infection.

In another embodiment, the use in medical therapy of a compound of anyone of the Formulas (I), (II-A), (II-B), (II-C), (III-A), (III-B),(III-C), (III-D), (III-E), (III-F), (III-G), (III-H), (IV-AA), (IV-AB),(IV-AC), (IV-AD), (IV-AE), (IV-AF), (IV-AG), (IV-AH), (IV-BA), (IV-BB),(IV-BC), (IV-BD), (IV-BE), (IV-BF), (IV-BG), and (IV-BH), as notedabove, or a pharmaceutical composition thereof, is provided.

In another embodiment, the use of a compound of any one of the Formulas(I), (II-A), (II-B), (II-C), (III-A), (III-B), (III-C), (III-D), (II-E),(III-F), (III-G), (III-H), (IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE),(IV-AF), (IV-AG), (IV-AH), (IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE),(IV-BF), (IV-BG), and (IV-BH), as noted above, or a pharmaceuticalcomposition thereof, for use in the therapeutic treatment of an HIVinfection is provided. In another embodiment, the use of a compound ofany one of the Formulas (I), (II-A), (II-B), (II-C), (III-A), (III-B),(III-C), (III-D), (III-E), (III-F), (III-G), (III-H), (IV-AA), (IV-AB),(IV-AC), (IV-AD), (IV-AE), (IV-AF), (IV-AG), (IV-AH), (IV-BA), (IV-BB),(IV-BC), (IV-BD), (IV-BE), (IV-BF), (IV-BG), and (IV-BH), as notedabove, or a pharmaceutical composition thereof, for use in theprophylactic or therapeutic treatment of an HIV infection is provided.

It is understood that any embodiment of the compounds of Formulas (I),(II-A), (II-B), (II-C), (III-A), (III-B), (III-C), (III-D), (III-E),(III-F), (III-G), (III-H), (IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE),(IV-AF), (IV-AG), (IV-AH), (IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE),(IV-BF), (IV-BG), and (IV-BH), as set forth above, and any specificsubstituent set forth herein for a R¹, R^(a), X, W, Y¹, Y², L, Z¹, Z²,Z³, or Z⁴ group in the compounds of Formulas (I), (II-A), (II-B),(II-C), (III-A), (III-B), (I-C), (III-D), (III-E), (III-F), (III-G),(III-H), (IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE), (IV-AF), (IV-AG),(IV-AH), (IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE), (IV-BF), (IV-BG),and (IV-BH), as set forth above, may be independently combined withother embodiments and/or substituents of compounds of Formulas (I),(II-A), (II-B), (II-C), (III-A), (III-B), (III-C), (II-D), (III-E),(III-F), (III-G), (III-H), (IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE),(IV-AF), (IV-AG), (IV-AH), (IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE),(IV-BF), (IV-BG), and (IV-BH), to form embodiments of the inventions notspecifically set forth above. In addition, in the event that a list ofsubstitutents is listed for any particular R¹, R^(a), X, W, Y¹, Y², L,Z¹, Z², Z³, or Z⁴ in a particular embodiment and/or claim, it isunderstood that each individual substituent may be deleted from theparticular embodiment and/or claim and that the remaining list ofsubstituents will be considered to be within the scope of the invention.

As one of skill in the art will appreciate, compounds of Formulas (I),(II-A), (II-B), (II-C), (IV-AA), (IV-AB), (IV-AC), (IV-AD), (IV-AE),(IV-AF), (IV-AG), (IV-AH), (IV-BA), (IV-BB), (IV-BC), (IV-BD), (IV-BE),(IV-BF), (IV-BG), and (IV-BH), wherein Z¹ and Z² or Z¹ and Z³, takentogether, form -L- may be shown in several different ways. For example,the Compound 3 of Example 3 may be shown as:

Pharmaceutical Compositions

For the purposes of administration, in certain embodiments, thecompounds described herein are administered as a raw chemical or areformulated as pharmaceutical compositions. Pharmaceutical compositionsdisclosed herein include a compound of Formula (I) and one or more of: apharmaceutically acceptable carrier, diluent or excipient. The compoundof Formula (I) is present in the composition in an amount which iseffective to treat a particular disease or condition of interest. Theactivity of compounds of Formula (I) can be determined by one skilled inthe art, for example, as described in the Examples below. Appropriateconcentrations and dosages can be readily determined by one skilled inthe art. In certain embodiments, a compound of Formula (I) is present inthe pharmaceutical composition in an amount from about 25 mg to about500 mg. In certain embodiments, a compound of Formula (I) is present inthe pharmaceutical composition in an amount of about 100 mg to about 300mg. In certain embodiments, a compound of Formula (I) is present in thepharmaceutical composition in an amount of about 25 mg, 50 mg, 100 mg,200 mg, 300 mg, 400 mg or about 500 mg.

Administration of the compounds of the invention, or theirpharmaceutically acceptable salts, in pure form or in an appropriatepharmaceutical composition, is carried out via any of the accepted modesof administration of agents for serving similar utilities. Thepharmaceutical compositions of the invention are prepared by combining acompound of the invention with an appropriate pharmaceuticallyacceptable carrier, diluent or excipient, and in specific embodimentsare formulated into preparations in solid, semi-solid, liquid or gaseousforms, such as tablets, capsules, powders, granules, ointments,solutions, suppositories, injections, inhalants, gels, microspheres, andaerosols. Exemplary routes of administering such pharmaceuticalcompositions include, without limitation, oral, topical, transdermal,inhalation, parenteral, sublingual, buccal, rectal, vaginal, andintranasal. Pharmaceutical compositions of the invention are formulatedso as to allow the active ingredients contained therein to bebioavailable upon administration of the composition to a patient.

Compositions that will be administered to a subject or patient take theform of one or more dosage units, where for example, a tablet may be asingle dosage unit, and a container of a compound of the invention inaerosol form may hold a plurality of dosage units. Actual methods ofpreparing such dosage forms are known, or will be apparent, to thoseskilled in this art; for example, see Remington: The Science andPractice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy andScience, 2000). The composition to be administered will, in any event,contain a therapeutically effective amount of a compound of theinvention, or a pharmaceutically acceptable salt thereof, for treatmentof a disease or condition of interest in accordance with the teachingsdescribed herein.

The pharmaceutical compositions disclosed herein are prepared bymethodologies well known in the pharmaceutical art. For example, incertain embodiments, a pharmaceutical composition intended to beadministered by injection is prepared by combining a compound of theinvention with sterile, distilled water so as to form a solution. Insome embodiments, a surfactant is added to facilitate the formation of ahomogeneous solution or suspension. Surfactants are compounds thatnon-covalently interact with the compound of the invention so as tofacilitate dissolution or homogeneous suspension of the compound in theaqueous delivery system.

The compounds of the invention, or their pharmaceutically acceptablesalts, are administered in a therapeutically effective amount, whichwill vary depending upon a variety of factors including the activity ofthe specific compound employed; the metabolic stability and length ofaction of the compound; the age, body weight, general health, sex, anddiet of the patient; the mode and time of administration; the rate ofexcretion; the drug combination; the severity of the particular disorderor condition; and the subject undergoing therapy.

Combination Therapy

In one embodiment, a method for treating or preventing an HIV infectionin a human having or at risk of having the infection is provided,comprising administering to the human a therapeutically effective amountof a compound disclosed herein, or a pharmaceutically acceptable saltthereof, in combination with a therapeutically effective amount of oneor more additional therapeutic agents.

In one embodiment, pharmaceutical compositions comprising a compounddisclosed herein, or a pharmaceutically acceptable salt thereof, incombination with one or more additional therapeutic agents, and apharmaceutically acceptable carrier, diluent or excipient are provided.

In one embodiment, combination pharmaceutical agents comprising acompound disclosed herein, or a pharmaceutically acceptable saltthereof, in combination with one or more additional therapeutic agentsare provided.

In the above embodiments, the additional therapeutic agent may be ananti-HIV agent. For example, in some embodiments, the additionaltherapeutic agent is selected from the group consisting of HIV proteaseinhibitors, HIV non-nucleoside inhibitors of reverse transcriptase, HIVnucleoside inhibitors of reverse transcriptase, HIV nucleotideinhibitors of reverse transcriptase, HIV integrase inhibitors, HIVnon-catalytic site (or allosteric) integrase inhibitors, entryinhibitors (e.g., CCR5 inhibitors, gp41 inhibitors (i.e., fusioninhibitors) and CD4 attachment inhibitors), CXCR4 inhibitors, gp120inhibitors, G6PD and NADH-oxidase inhibitors, compounds that target theHIV capsid (“capsid inhibitors”; e.g., capsid polymerization inhibitorsor capsid disrupting compounds such as those disclosed in WO 2013/006738(Gilead Sciences), US 2013/0165489 (University of Pennsylvania), and WO2013/006792 (Pharma Resources), pharmacokinetic enhancers, and otherdrugs for treating HIV, and combinations thereof. In furtherembodiments, the additional therapeutic agent is selected from one ormore of:

(1) HIV protease inhibitors selected from the group consisting ofamprenavir, atazanavir, fosamprenavir, indinavir, lopinavir, ritonavir,nelfinavir, saquinavir, tipranavir, brecanavir, darunavir, TMC-126,TMC-114, mozenavir (DMP-450), JE-2147 (AG1776), L-756423, RO0334649,KNI-272, DPC-681, DPC-684, GW640385X, DG17, PPL-100, DG35, and AG 1859;

(2) HIV non-nucleoside or non-nucleotide inhibitors of reversetranscriptase selected from the group consisting of capravirine,emivirine, delaviridine, efavirenz, nevirapine, (+) calanolide A,etravirine, GW5634, DPC-083, DPC-961, DPC-963, MIV-150, TMC-120,rilpivirene, BILR 355 BS, VRX 840773, lersivirine (UK-453061), RDEA806,KM023 and MK-1439;

(3) HIV nucleoside inhibitors of reverse transcriptase selected from thegroup consisting of zidovudine, emtricitabine, didanosine, stavudine,zalcitabine, lamivudine, abacavir, amdoxovir, elvucitabine, alovudine,MIV-210, ±-FTC, D-d4FC, emtricitabine, phosphazide, fozivudine tidoxil,apricitibine (AVX754), KP-1461, GS-9131 (Gilead Sciences) andfosalvudine tidoxil (formerly HDP 99.0003);

(4) HIV nucleotide inhibitors of reverse transcriptase selected from thegroup consisting of tenofovir, tenofovir disoproxil fumarate, tenofoviralafenamide fumarate (Gilead Sciences), GS-7340 (Gilead Sciences),GS-9148 (Gilead Sciences), adefovir, adefovir dipivoxil, CMX-001(Chimerix) or CMX-157 (Chimerix);

(5) HIV integrase inhibitors selected from the group consisting ofcurcumin, derivatives of curcumin, chicoric acid, derivatives ofchicoric acid, 3,5-dicaffeoylquinic acid, derivatives of3,5-dicaffeoylquinic acid, aurintricarboxylic acid, derivatives ofaurintricarboxylic acid, caffeic acid phenethyl ester, derivatives ofcaffeic acid phenethyl ester, tyrphostin, derivatives of tyrphostin,quercetin, derivatives of quercetin, S-1360, AR-177, L-870812, andL-870810, raltegravir, BMS-538158, GSK364735C, BMS-707035, MK-2048, BA011, elvitegravir, dolutegravir and GSK-744;

(6) HIV non-catalytic site, or allosteric, integrase inhibitors (NCINI)including, but not limited to, BI-224436, CX0516, CX05045, CX14442,compounds disclosed in WO 2009/062285 (Boehringer Ingelheim), WO2010/130034 (Boehringer Ingelheim), WO 2013/159064 (Gilead Sciences), WO2012/145728 (Gilead Sciences), WO 2012/003497 (Gilead Sciences), WO2012/003498 (Gilead Sciences) each of which is incorporated byreferences in its entirety herein;

(7) gp41 inhibitors selected from the group consisting of enfuvirtide,sifuvirtide, albuvirtide, FB006M, and TRI-1144;

(8) the CXCR4 inhibitor AMD-070;

(9) the entry inhibitor SP01A:

(10) the gp120 inhibitor BMS-488043;

(11) the G6PD and NADH-oxidase inhibitor immunitin;

(12) CCR5 inhibitors selected from the group consisting of aplaviroc,vicriviroc, maraviroc, cenicriviroc, PRO-140, INCB15050, PF-232798(Pfizer), and CCR5mAb004;

(13) CD4 attachment inhibitors selected from the group consisting ofibalizumab (TMB-355) and BMS-068 (BMS-663068);

(14) pharmacokinetic enhancers selected from the group consisting ofcobicistat and SPI-452; and

(15) other drugs for treating HIV selected from the group consisting ofBAS-100, SPI-452, REP 9, SP-01A, TNX-355, DES6, ODN-93, ODN-112, VGV-1,PA-457 (bevirimat), HRG214, VGX-410, KD-247, AMZ 0026, CYT 99007A-221HIV, DEBIO-025, BAY 50-4798, MDXO10 (ipilimumab), PBS 119, ALG 889, andPA-1050040 (PA-040),

and combinations thereof

In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with two, three,four or more additional therapeutic agents. In certain embodiments, acompound disclosed herein, or a pharmaceutically acceptable saltthereof, is combined with two additional therapeutic agents. In otherembodiments, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with three additional therapeuticagents. In further embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with fouradditional therapeutic agents. The two, three four or more additionaltherapeutic agents can be different therapeutic agents selected from thesame class of therapeutic agents, or they can be selected from differentclasses of therapeutic agents. In a specific embodiment, a compounddisclosed herein, or a pharmaceutically acceptable salt thereof, iscombined with an HIV nucleotide inhibitor of reverse transcriptase andan HIV non-nucleoside inhibitor of reverse transcriptase. In anotherspecific embodiment, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with an HIV nucleotide inhibitor ofreverse transcriptase, and an HIV protease inhibiting compound. In afurther embodiment, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with an HIV nucleotide inhibitor ofreverse transcriptase, an HIV non-nucleoside inhibitor of reversetranscriptase, and an HIV protease inhibiting compound. In an additionalembodiment, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with an HIV nucleotide inhibitor ofreverse transcriptase, an HIV non-nucleoside inhibitor of reversetranscriptase, and a pharmacokinetic enhancer.

In certain embodiments, when a compound disclosed herein is combinedwith one or more additional therapeutic agents as described above, thecomponents of the composition are administered as a simultaneous orsequential regimen. When administered sequentially, the combination maybe administered in two or more administrations.

In certain embodiments, a compound disclosed herein is combined with oneor more additional therapeutic agents in a unitary dosage form forsimultaneous administration to a patient, for example as a solid dosageform for oral administration.

In certain embodiments, a compound disclosed herein is administered withone or more additional therapeutic agents. Co-administration of acompound disclosed herein with one or more additional therapeutic agentsgenerally refers to simultaneous or sequential administration of acompound disclosed herein and one or more additional therapeutic agents,such that therapeutically effective amounts of the compound disclosedherein and one or more additional therapeutic agents are both present inthe body of the patient.

Co-administration includes administration of unit dosages of thecompounds disclosed herein before or after administration of unitdosages of one or more additional therapeutic agents, for example,administration of the compound disclosed herein within seconds, minutes,or hours of the administration of one or more additional therapeuticagents. For example, in some embodiments, a unit dose of a compounddisclosed herein is administered first, followed within seconds orminutes by administration of a unit dose of one or more additionaltherapeutic agents. Alternatively, in other embodiments, a unit dose ofone or more additional therapeutic agents is administered first,followed by administration of a unit dose of a compound disclosed hereinwithin seconds or minutes. In some embodiments, a unit dose of acompound disclosed herein is administered first, followed, after aperiod of hours (e.g., 1-12 hours), by administration of a unit dose ofone or more additional therapeutic agents. In other embodiments, a unitdose of one or more additional therapeutic agents is administered first,followed, after a period of hours (e.g., 1-12 hours), by administrationof a unit dose of a compound disclosed herein.

The following Examples illustrate various methods of making compounds ofthis invention, i.e., compound of Formula (I):

wherein R¹, X, W, Y¹, Y², Z¹, Z², or Z⁴ are as defined above. It isunderstood that one skilled in the art may be able to make thesecompounds by similar methods or by combining other methods known to oneskilled in the art. It is also understood that one skilled in the artwould be able to make, in a similar manner as described below, othercompounds of Formula (I) not specifically illustrated below by using theappropriate starting components and modifying the parameters of thesynthesis as needed. In general, starting components may be obtainedfrom sources such as Sigma Aldrich, Lancaster Synthesis, Inc.,Maybridge, Matrix Scientific, TCI, and Fluorochem USA, etc. orsynthesized according to sources known to those skilled in the art (see,for example, Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, 5th edition (Wiley, December 2000)) or prepared as describedherein.

The following examples are provided for purposes of illustration, notlimitation.

EXAMPLES General Synthetic Schemes

Schemes 1-3 are provided as further embodiments of the invention andillustrate general methods which were used to prepare compounds havingFormula (I) and which can be used to prepare additional compound havingFormula (I).

A1 can be converted to amide A2 with an appropriate amine and a couplingreagent such as HATU or EDCI. A2 can be converted to A3 with a strongacid such as methanesulfonic acid. A3 can be converted to either A5 orA4 by heating with an appropriate cyclic diamine or cyclic aminoalcoholfollowed by methyl deprotection with a reagent such as magnesiumbromide.

Alternatively, A1 can be converted to A6 by treatment with a strong acidsuch as methanesulfonic acid. A6 can be condensed with an appropriatecyclic diamine or cyclic aminoalcohol followed by methyl deprotectionwith a reagent such as magnesium bromide to form either A7 or A8respectively. A7 or A8 can be converted into amides A5 and A4 bytreatment with an appropriate amine and a coupling reagent such as HATUor EDCI followed by methyl deprotection with a reagent such as magnesiumbromide.

B1 (as described in WO2012/018065) is condensed with diamine underreflux condition to give B2. B2 is hydrolyzed and coupled with an amineby an amide-forming method to afford product B3 upon removal of a benzylprotecting group.

Representative Compounds Example 1 Preparation of Compound 1N-(2,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

1-(2,2-dimethoxyethyl)-5-methoxy-6-(methoxycarbonyl)-4-oxo-1,4-dihydropyridine-3-carboxylicacid (1-A, 0.300 g, 0.95 mmol), prepared as described in WO2011/119566A1, was evaporated once from dry toluene, suspended in acetonitrile (4mL) and treated with N,N-diisopropylethylamine (DIPEA) (0.329 mL, 1.90mmol), 2,4-difluorobenzylamine (0.125 mL, 1.05 mmol) and HATU (0.433 g,1.14 mmol). The reaction mixture was stirred for 10 minutes andconcentrated. The residue was purified by flash chromatography on silicagel (10 to 60% ethyl acetate:dichloromethane) to afford the compoundmethyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dimethoxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate,1-B. ¹H-NMR (400 MHz, DMSO-d6) δ 10.28 (t, J=6.0 Hz, 1H), 8.46 (s, 1H),7.42 (dd, J=15.4, 8.6 Hz, 1H), 7.24 (m, 1H), 7.06 (m, 1H), 4.52 (m, 3H),4.22 (d, J=4.4 Hz, 2H), 3.92 (s, 3H), 3.80 (s, 3H), 3.29 (d, 6H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₀H₂₃F₂N₂O₇: 441.15; found:441.2.

Step 2

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dimethoxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-B, 0.106 g, 0.24 mmol) in acetonitrile (0.9 mL) and acetic acid (0.1mL) was treated with methanesulfonic acid (0.005 mL, 0.072 mmol), sealedwith a yellow cap, and heated to 70° C. After 16 hours, the mixture wascooled to afford a crude solution of methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate,1-C. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₈H₁₉F₂N₂O₇: 413.12; found:413.1.

Steps 3 and 4

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 0.65 mL of the crude mixture from the previous step, 0.17 mmol)was treated with acetonitrile (0.65 mL) and cis-3-aminocyclpentanol(0.06 mL). The reaction mixture was sealed and heated to 90° C. After 30minutes, the reaction mixture was cooled and magnesium bromide (0.063 g,0.34 mmol) was added. The mixture was resealed and heated to 50° C.After 10 minutes, the reaction mixture was partitioned betweendichloromethane and hydrochloric acid (0.2 M aq). The organic layer wasremoved and the aqueous layer extracted again with dichlormethane. Thecombined organic layers were dried over sodium sulfate, filtered andconcentrated. Prep-HPLC purification (30-70% acetonitrile:water, 0.1%TFA) afforded Compound 1 as a racemic mixture. ¹H-NMR (400 MHz, DMSO-d6)δ 12.45 (br s, 1H), 10.35 (t, J=5.8 Hz, 1H), 8.45 (s, 1H), 7.37 (dd,J=15.4, 8.6 Hz, 1H), 7.23 (dt, J=2.5, 9.9 Hz, 1H), 7.05 (dt, J=2.2, 8.7Hz, 1H), 5.43 (dd, J=9.6, 4.0 Hz, 1H), 5.09 (br s, 1H), 4.68 (dd,J=13.2, 4.0 Hz, 1H), 4.59 (br s, 1H), 4.53 (m, 2H), 4.02 (dd, J=12.6,9.4 Hz), 1.93 (br s, 4H), 1.83 (d, J=12.0 Hz), 1.57 (dt, J=12.2, 3.2Hz). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₅: 432.14; found:432.2.

Examples 2 and 3 Preparation of Compounds 2 and 3(2R,5S,13aR)—N-(2,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide(2) and(2S,5R,13aS)—N-(2,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide(3)

Compound 1 (16 mg) was separated by chiral HPLC using Chiralpak AS-Hwith 100% ethanol as eluent to afford Compounds 2 and 3 inenantiomerically enriched form. For Compound 2: LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₁H₂₀F₂N₃O₅: 432.14; found: 432.2, Chiral HPLC retentiontime=4.50 minutes (Chiralpak AS-H, 150×4.6 mm, 1 mL/min EtOH). ForCompound 3: LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₅: 432.14;found: 432.2, Chiral HPLC retention time=6.84 minutes (Chiralpak AS-H,150×4.6 mm, 1 mL/min EtOH). ¹H-NMR (400 MHz, DMSO-d6) δ 12.45 (br s,1H), 10.35 (t, J=5.8 Hz, 1H), 8.44 (s, 1H), 7.37 (dd, J=15.2, 8.4 Hz,1H), 7.23 (m, 1H), 7.05 (dt, J=1.8 Hz, 8.7 Hz, 1H), 5.44 (dd, J=9.6, 4.0Hz), 5.09 (br s, 1H), 4.68 (dd, J=12.8, 4.0 Hz, 1H), 4.59 (br s, 1H),4.53 (m, 2H), 4.02 (dd, J=12.6, 9.4 Hz, 1H), 1.93 (br s, 4H), 1.83 (d,J=12.4 Hz, 1H), 1.57 (m, 1H).

Alternatively, Compound 3 was prepared as follows:

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 1.2 mmol in 5 mL of 9:1 acetonitrile:acetic acid containing 0.026mL methanesulfonic acid) was treated with acetonitrile (5.0 mL) andcis-3-aminocyclpentanol (0.24 g, 2.4 mmol). The reaction mixture wassealed and heated to 90° C. After 30 minutes, the reaction mixture wascooled, treated with potassium carbonate (0.332 g, 2.4 mmol), sealed andreheated to 90° C. After 15 minutes, the mixture was cooled andpartitioned between dichlormethane and hydrochloric acid (0.2 Maqueous). The organic layer was removed and the aqueous solution wasextracted again with dichloromethane. The combined organic layers weredried over sodium sulfate (anhydrous), filtered and concentrated. Theresidue was purified by flash chromatography (0-8% ethanol (containing11% saturated aqueous ammonium hydroxide) in dichloromethane) to affordIntermediate 1-D. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₂₂F₂N₃O₅:446.15; found: 446.2.

Intermediate 1-D (270 mg) was separated by chiral SFC on a 50 mmChiralpak AD-H column using 50% (1:1 methanol:acetonitrile) insupercritical carbon dioxide as eluent to afford Intermediates 3-A(first eluting peak) and 3-B (second eluting peak) in enantioenrichedform. For 3-A: LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₂₂F₂N₃O₅:446.15; found: 446.2. For 3-B: LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₂F₂N₃O₅: 446.15; found: 446.2.

Intermediate 3-A (0.110 g, 0.247 mmol) in acetonitrile (5 mL) wastreated portion wise with magnesium bromide (0.091 g, 0.494 mmol),sealed and heated to 50° C. After 10 minutes the mixture was cooled andpartitioned between dichloromethane and hydrochloric acid (0.2 Maqueous). The organic layer was separated and the aqueous extractedagain with dichloromethane. The combined organic layers were dried oversodium sulfate, filtered and concentrated. Preparative HPLC purification(30-70% acetonitrile:water, 0.1% TFA) afforded Compound 3 inenantioenriched form. Chiral HPLC retention time=6.51 minutes (ChiralpakAS-H, 150×4.6 mm, 1 mL/min EtOH). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₂₀F₂N₃O₅: 432.14; found: 432.2. ¹H-NMR (400 MHz, DMSO-d6) δ 12.45(br s, 1H), 10.35 (t, J=5.8 Hz, 1H), 8.44 (s, 1H), 7.37 (dd, J=15.2, 8.4Hz, 1H), 7.23 (m, 1H), 7.05 (dt, J=1.8 Hz, 8.7 Hz, 1H), 5.44 (dd, J=9.6,4.0 Hz), 5.09 (br s, 1H), 4.68 (dd, J=12.8, 4.0 Hz, 1H), 4.59 (br s,1H), 4.53 (m, 2H), 4.02 (dd, J=12.6, 9.4 Hz, 1H), 1.93 (br s, 4H), 1.83(d, J=12.4 Hz, 1H), 1.57 (m, 1H).

Example 4 Preparation of Compound 4(1S,4R)—N-(2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-1,4-methanopyrido[1′,2′:4,5]pyrazino[1,2-a]pyrimidine-9-carboxamide

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 0.12 mmol in 0.53 mL of 9:1 acetonitrile:acetic acid containing0.002 mL methanesulfonic acid) was treated with acetonitrile then(R)-pyrrolidin-3-amine (0.032 mL, 0.36 mmol). The reaction mixture wascapped and heated to 90° C. for 5.5 hours. After cooling, the mixturewas partitioned between dichloromethane and sodium bicarbonate (1Maqueous). The organic layer was separated and the aqueous was extractedagain with ethyl acetate. The combined organic layers were dried oversodium sulfate (anhydrous), filtered and concentrated. The residue wasdissolved in acetonitrile (1 mL), treated with magnesium bromide (0.022g, 0.12 mmol), capped and heated to 50° C. for 10 minutes. After coolingthe mixture was partitioned between dichloromethane and ammoniumchloride (sat). The organic layer was separated and the aqueous wasextracted again with dichloromethane. The aqueous layer was adjusted topH=1 with HCl (aq) and extracted again with dichloromethane. The aqueoussolution was adjusted to pH=3 with NaOH (aq) and extracted again withdichloromethane. The combined organic layers were dried over sodiumsulfate, filtered, and concentrated. Preparative HPLC purification(10-55% acetonitrile:water, 0.1% TFA) afforded Compound 4. ¹H-NMR (400MHz, CD₃OD-d4) δ 8.42 (s, 1H), 7.42, (q, J=7.7 Hz, 1H), 6.99-6.90 (m,2H), 5.07 (br s, 1H), 4.73 (br d, J=10.8 Hz, 1H), 4.62 (s, 2H), 4.51 (brd, J=12.8 Hz, 1H), 4.07 (t, J=11.8 Hz, 1H), 3.4-3.0 (m, 3H), 2.76 (br d,J=8.8 Hz, 1H), 2.15-2.0 (m, 1H), 1.9-1.8 (m, 1H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₀H₁₉F₂N₄O₄: 417.14; found: 417.2.

Example 5 Preparation of Compound 5(4R,12aS)—N-(1-(2,4-difluorophenyl)cyclopropyl)-7-hydroxy-4-methyl-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-[1,3]oxazino[3,2-d]pyrido[1,2-a]pyrazine-9-carboxamide

Step 1

(4R,12aS)-7-methoxy-4-methyl-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-[1,3]oxazino[3,2-d]pyrido[1,2-a]pyrazine-9-carboxylicacid (Intermediate 5-A) was prepared in an analogous manner to(3S,11aR)-6-methoxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydrooxazolo[3,2-d]pyrido[1,2-a]pyrazine-8-carboxylicacid as described in WO2011/119566, substituting (R)-3-aminobutan-1-olfor (S)-2-aminopropan-1-ol. WO2011/119566 is incorporated herein byreference in its entirety. A suspension of Intermediate 5-A (24.8 mg,0.080 mmol), 1-(2,4-difluorophenyl)cyclopropanamine HCl salt (5-B, 21.9mg, 0.107 mmol), and HATU (48 mg, 0.126 mmol) in CH₂Cl₂ (2 mL) wasstirred at ambient temperature as N,N-diisopropylethylamine (DIPEA) (0.1mL, 0.574 mmol) was added. After 30 minutes, the reaction mixture wasdiluted with ethyl acetate before washing with 10/o aqueous citric acidsolution (×1) and saturated aqueous NaHCO₃ solution (×1). After theaqueous fractions were extracted with ethyl acetate (×1), the organicfractions were combined, dried (MgSO₄), and concentrated. The residuewas purified by combiflash (12 g column) using hexanes, ethyl acetate,and 200% methanol in ethyl acetate to obtain(4R,12aS)—N-(1-(2,4-difluorophenyl)cyclopropyl)-7-methoxy-4-methyl-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-[1,3]oxazino[3,2-d]pyrido[1,2-a]pyrazine-9-carboxamide,Intermediate 5-C. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₃H₂₄F₂N₃O₅:460.17; found 460.2.

Step 2

A suspension of Intermediate 5-C (39 mg, 0.080 mmol) and magnesiumbromide (42 mg, 0.2282 mmol) in acetonitrile (2 mL) was stirred at 50°C. After 1 hour, the reaction mixture was stirred at 0° C. bath when 1 NHCl (2 mL) was added. After the resulting mixture was diluted with water(˜20 mL), the product was extracted with dichloromethane (×3) and thecombined extracts were dried (MgSO₄) and concentrated. The residue waspurified by preparative HPLC to obtain(4R,12aS)—N-(1-(2,4-difluorophenyl)cyclopropyl)-7-hydroxy-4-methyl-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-[1,3]oxazino[3,2-d]pyrido[1,2-a]pyrazine-9-carboxamide,compound 5, as TFA salt. ¹H-NMR (400 MHz, CDCl₃) δ 10.72 (br s, 1H),8.37 (s, 1H), 7.57 (d, J=7.9 Hz, 1H), 6.71-6.81 (m, 2H), 5.23 (dd, J=5.6and 4.4 Hz, 1H), 4.98 (br quint, J=˜6.5 Hz, 1H), 4.26 (dd, J=13.6 and4.4 Hz, 1H), 4.12 (dd, J=13.6 and 5.6 Hz, 1H), 4.00-4.06 (m, 2H),2.16-2.25 (m, 1H), 1.55 (br dd, J=13.8 and 1.8 Hz, 1H), 1.40 (d, J=6.8Hz, 3H), 1.22-1.31 (m, 4H). ¹⁹F NMR (376.1 MHz, CDCl₃) δ −76.38 (s, 3F),−111.69˜−111.645 (m, 2F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₂F₂N₃O₅: 446.15; found: 446.2.

Example 6 Preparation of Compound 6(1R,4S)—N-(2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-1,4-methanopyrido[1′,2′:4,5]pyrazino[1,2-a]pyrimidine-9-carboxamide

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 0.100 g, 0.243 mmol), (S)-pyrrolidin-3-amine (0.043 mL, 0.485mmol) and potassium carbonate (0.067 g, 0.485 mmol) were suspended inacetonitrile (1.9 mL) and acetic acid (0.1 mL) and heated to 90° C. for1.5 hours. After cooling, the mixture was treated with magnesium bromide(0.090 g) and heated to 50° C. for 30 minutes. After cooling, themixture partitioned between dichloromethane and 0.2 M HCl. The organiclayer was separated and the aqueous was extracted again withdichloromethane. The combined organic layers were dried over sodiumsulfate (anhydrous), filtered and concentrated. Preparative HPLCpurification (25-50% acetonitrile:water, 0.1% TFA) afforded Compound 6.¹H-NMR (400 MHz, DMSO-d₆) δ 10.33 (t, J=6.0 Hz, 1H), 8.44 (s, 1H),7.48-7.32 (m, 1H), 7.31-7.15 (m, 1H), 7.14-6.97 (m, 1H), 4.86 (d, J=2.9Hz, 1H), 4.62-4.54 (m, 1H), 4.52 (d, J=5.9 Hz, 1H), 4.01 (d, J=13.0 Hz,1H), 2.99-2.76 (m, 3H), 1.96-1.81 (m, 1H), 1.71-1.53 (m, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₀H₁₉F₂N₄O₄: 417.14; found: 417.2.

Example 7 Preparation of Compound 7(2S,6R)—N-(2,4-difluorobenzyl)-9-hydroxy-8,10-dioxo-3,4,5,6,8,10,14,14a-octahydro-2H-2,6-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazocine-11-carboxamide

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 0.050 g, 0.121 mmol), (1S,3R)-3-aminocyclohexanol (0.028 g, 0.243mmol) and potassium carbonate (0.034 g, 0.243 mmol) were suspended inacetonitrile (0.95 mL) and heated to 90° C. for 0.5 hour. After cooling,acetic acid (0.050 mL) was added and the mixture was reheated to 90° C.for 2 h. After cooling the mixture was treated with magnesium bromide(0.044 g) and heated to 50° C. for 1 hour. After cooling, a secondportion of magnesium bromide (0.044 g) was added and the mixture wasreheated to 50° C. for 15 minutes. After cooling, the mixturepartitioned between dichloromethane and 0.2 M HCl. The organic layer wasseparated and the aqueous was extracted again with dichloromethane. Thecombined organic layers were dried over sodium sulfate (anhydrous),filtered and concentrated. Preparative HPLC purification (40-80%acetonitrile:water, 0.1% TFA) afforded Compound 7. ¹H-NMR (400 MHz,DMSO-d₆) δ 12.40 (s, 1H), 10.36 (t, J=6.1 Hz, 1H), 8.45 (s, 1H),7.48-7.29 (m, 1H), 7.31-7.13 (m, 1H), 7.13-6.97 (m, 1H), 5.56 (dd,J=10.0, 4.1 Hz, 1H), 4.70 (dd, J=12.7, 4.1 Hz, 1H), 4.52 (d, J=5.5 Hz,2H), 4.40-4.29 (m, 2H), 4.06 (dd, J=12.5, 10.2 Hz, 1H), 2.46-2.36 (m,1H), 1.98-1.63 (m, 4H), 1.57-1.30 (m, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₂H₂₂F₂N₃O₅: 446.15; found: 446.2.

Example 8 Preparation of Compound 8(2R,6S)—N-(2,4-difluorobenzyl)-9-hydroxy-8,10-dioxo-3,4,5,6,8,10,14,14a-octahydro-2H-2,6-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazocine-11-carboxamide

Compound 8 was prepared in a similar manner to compound 7 using(1R,3S)-3-aminocyclohexanol in place of (1S,3R)-3-aminocyclohexanol.¹H-NMR (400 MHz, DMSO-d₆) δ 12.40 (s, 1H), 10.36 (t, J=6.1 Hz, 1H), 8.45(s, 1H), 7.48-7.30 (m, 1H), 7.23 (td, J=10.6, 2.7 Hz, 1H), 7.05 (td,J=8.3, 2.3 Hz, 1H), 5.56 (dd, J=10.1, 4.1 Hz, 1H), 4.70 (dd, J=12.8, 3.9Hz, 1H), 4.52 (d, J=5.6 Hz, 2H), 4.39-4.27 (m, 2H), 4.06 (dd, J=12.6,10.0 Hz, 1H), 2.47-2.35 (m, 1H), 2.00-1.64 (m, 4H), 1.58-1.30 (m, 3H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₂₂F₂N₃O₅: 446.15; found:446.2.

Examples 9 and 10 Preparation of Compounds 9 and 10(2S,5R,13aS)—N—((R)-1-(4-fluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide9 and(2R,5S,13aR)—N—((R)-1-(4-fluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide10

Step 1

1-(2,2-dimethoxyethyl)-5-methoxy-6-(methoxycarbonyl)-4-oxo-1,4-dihydropyridine-3-carboxylicacid (1-A, 0.500 g, 1.59 mmol), was suspended in acetonitrile (6 mL) andtreated with N,N-diisopropylethylamine (DIPEA) (0.550 mL, 3.17 mmol),(R)-1-(4-fluorophenyl)ethanamine (0.242 mg, 1.74 mmol) and HATU (0.661g, 1.74 mmol). The reaction mixture was stirred for 2 hours andpartitioned between ethyl acetate and water. The organic layer wasseparated and washed with HCl (10% aq), sodium bicarbonate (1M aq),dried over sodium sulfate, filtered and concentrated to afford crude(R)-methyl1-(2,2-dimethoxyethyl)-5-(1-(4-fluorophenyl)ethylcarbamoyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylatewhich was used without purification in the next step: LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₁H₂₆FN₂O₇: 437.17; found: 437.1.

Step 2

(R)-methyl1-(2,2-dimethoxyethyl)-5-(1-(4-fluorophenyl)ethylcarbamoyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylatewas suspended in acetonitrile (5.7 mL) and acetic acid (0.6 mL) andtreated with methane sulfonic acid (0.031 mL, 0.477 mmol). The mixturewas capped and heated to 75° C. After 7 h, the mixture was cooled andused without purification in the next step: LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₁₉H₂₂FN₂O₇: 409.14; found: 409.0.

Step 3

(R)-methyl1-(2,2-dihydroxyethyl)-5-(1-(4-fluorophenyl)ethylcarbamoyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(3.6 mL of the crude mixture from Step 2, 0.8 mmol) was diluted withacetonitrile (3.6 mL) and treated with cis-3-aminocyclpentanol, HCl salt(0.219 g, 1.6 mmol) and potassium carbonate (0.276 g, 2.0 mmol). Themixture was capped and heated to 90° C. After 20 minutes, the reactionmixture was cooled and partitioned between dichloromethane and HCl (0.2M aq). The layers were separated and the aqueous layer was extractedagain with dichloromethane. The combined organic layers were treatedwith a small amount of acetonitrile, dried over sodium sulfate, filteredand concentrated.

The residue was suspended in acetonitrile (4 mL) and treated withmagnesium bromide (0.177 g). The mixture was capped and heated to 50° C.After 10 minutes, the reaction mixture was cooled and partitionedbetween dichloromethane and HCl (0.2 M aq). The layers were separatedand the aqueous layer was extracted again with dichlormethane. Thecombined organic layers were dried over sodium sulfate, filtered andconcentrated. The residue was purified by flash chromatography on silicagel (0-8% ethanol:DCM) to afford a diastereomeric mixture of desired 9and 10.

The mixture was separated by chiral HPLC using Chiralpak AD-H with 100%ethanol as eluent to afford Compounds 9 and 10 in enantiomericallyenriched form:

For Compound 9: LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₂₃FN₃O₅:428.16; found: 428.1. Chiral HPLC retention time=10.177 minutes(Chiralpak AD-H, 150×4.6 mm, 1 mL/min EtOH). ¹H-NMR (400 MHz, DMSO-d₆) δ12.45 (s, 1H), 10.45 (d, J=7.7 Hz, 1H), 8.40 (s, 1H), 7.37 (dd, J=8.6,5.6 Hz, 2H), 7.15 (t, J=8.9 Hz, 2H), 5.44 (dd, J=9.5, 4.2 Hz, 1H),5.17-5.04 (m, 2H), 4.73-4.62 (m, 1H), 4.59 (s, 1H), 4.00 (dd, J=12.7,9.5 Hz, 1H), 1.93 (s, 4H), 1.83 (d, J=11.8 Hz, 1H), 1.56 (dt, J=12.1,3.4 Hz, 1H), 1.44 (d, J=6.9 Hz, 3H).

For Compound 10: LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₂₃FN₃O₅:428.16; found: 428.1. Chiral HPLC retention time=14.061 minutes(Chiralpak AD-H, 150×4.6 mm, 1 mL/min EtOH). ¹H-NMR (400 MHz, DMSO-d₆) δ12.44 (s, 1H), 10.46 (d, J=7.8 Hz, 1H), 8.41 (s, 1H), 7.37 (dd, J=8.6,5.6 Hz, 2H), 7.15 (t, J=8.9 Hz, 2H), 5.42 (dd, J=9.6, 4.1 Hz, 1H),5.18-5.02 (m, 2H), 4.67 (dd, J=12.8, 4.2 Hz, 1H), 4.59 (s, 1H), 4.02(dd, J=12.7, 9.6 Hz, 1H), 1.93 (s, 4H), 1.83 (d, J=12.0 Hz, 1H), 1.57(dt, J=13.0, 3.5 Hz, 1H), 1.44 (d, J=6.9 Hz, 3H).

Example 11 Preparation of Compound 11(2S,5R,13aS)—N—((R)-1-(2,4-difluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

1-(2,2-dimethoxyethyl)-5-methoxy-6-(methoxycarbonyl)-4-oxo-1,4-dihydropyridine-3-carboxylicacid (1-A, 0.315 g, 1.00 mmol), was suspended in acetonitrile (4 mL) andtreated with N,N-diisopropylethylamine (DIPEA) (0.348 mL, 2.00 mmol),(R)-1-(2,4-difluorophenyl)ethanamine HCl salt (0.213 mg, 1.10 mmol) andHATU (0.418 g, 1.10 mmol). The reaction mixture was stirred for 1 hourand partitioned between dichloromethane and HCl (10% aq). The organiclayer was separated and washed sodium bicarbonate (1M aq), dried oversodium sulfate, filtered and concentrated to afford crude (R)-methyl5-(1-(2,4-difluorophenyl)ethylcarbamoyl)-1-(2,2-dimethoxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylatewhich was used without purification in the next step. LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₁H₂₅F₂N₂O₇: 455.16; found: 455.1.

Step 2

(R)-methyl5-(1-(2,4-difluorophenyl)ethylcarbamoyl)-1-(2,2-dimethoxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylatewas suspended in acetonitrile (3.6 mL) and acetic acid (0.4 mL) andtreated with methane sulfonic acid (0.020 mL). The mixture was cappedand heated to 75° C. After 16 hours, the crude mixture was cooled andused without purification in the next step. LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₁₉H₂₁F₂N₂O₇: 427.13; found: 427.1.

Step 3

(R)-methyl5-(1-(2,4-difluorophenyl)ethylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(half of the crude mixture from Step 2, approx 0.5 mmol) was dilutedwith acetonitrile (2.5 mL) and treated with (1S,3R)-3-aminocyclopentanol(0.110 g, 1.09 mmol) and potassium carbonate (0.069 g, 0.50 mmol). Themixture was capped and heated to 90° C. After 15 minutes, the reactionmixture was cooled and magnesium bromide (0.184 g) was added. Thereaction mixture was heated to 50° C. After 10 minutes, the mixture wascooled and treated with an additional portion of magnesium bromide(0.184 g). The reaction mixture was reheated to 50° C. and stirred for10 minutes. After cooling, the mixture was partitioned betweendichloromethane and HCl (0.2 M aq). The layers were separated and theaqueous layer was extracted again with dichloromethane. The combinedorganic layers were dried over sodium sulfate, filtered andconcentrated. Preparative HPLC purification (30-60%/oacetonitrile:water, 0.1% TFA) afforded desired Compound 11. LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₂H₂₂F₂N₃O₅: 446.15; found: 446.1. ¹H-NMR(400 MHz, DMSO-d) δ 12.46 (s, 1H), 10.53 (d, J=7.5 Hz, 1H), 8.38 (s,1H), 7.39 (q, J=8.5 Hz, 1H), 7.29-7.12 (m, 1H), 7.13-6.93 (m, 1H), 5.44(dd, J=9.8, 4.2 Hz, 1H), 5.28 (p, J=7.3, 6.8 Hz, 1H), 5.09 (s, 1H), 4.66(dd, J=13.2, 4.3 Hz, 1H), 4.59 (s, 1H), 3.99 (dd, J=13.1, 9.6 Hz, 1H),1.93 (s, 4H), 1.83 (d, J=12.4 Hz, 1H), 1.56 (dt, J=12.5, 2.9 Hz, 1H),1.45 (d, J=6.9 Hz, 3H).

Example 12 Preparation of Compound 12(2R,5S,13aR)—N—((R)-1-(2,4-difluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 12 was prepared in a similar manner to compound 11 using(1R,3S)-3-aminocyclopentanol in place of (1S,3R)-3-aminocyclopentanol.¹H-NMR (400 MHz, DMSO-d₆) δ 12.43 (s, 1H), 10.52 (d, J=8.2 Hz, 1H), 8.38(s, 1H), 7.39 (q, J=8.4 Hz, 1H), 7.28-7.12 (m, 1H), 7.11-6.97 (m, 1H),5.41 (dd, J=10.0, 4.0 Hz, 1H), 5.35-5.20 (m, 1H), 5.08 (s, 1H), 4.65(dd, J=13.1, 3.8 Hz, 1H), 4.58 (s, 1H), 4.01 (dd, J=12.8, 9.5 Hz, 1H),1.92 (s, 4H), 1.83 (d, J=11.5 Hz, 1H), 1.61-1.51 (m, 1H), 1.44 (d, J=6.9Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₂₂F₂N₃O₅: 446.15;found: 446.1.

Example 13 Preparation of Compound 13(2S,5R,13aS)—N—((S)-1-(2,4-difluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 13 was prepared in a similar manner to compound 11 using(S)-1-(2,4-difluorophenyl)ethanaminein place of(R)-1-(2,4-difluorophenyl)ethanamine, and using only a single portion ofmagnesium bromide (0.184 g). ¹H-NMR (400 MHz, DMSO-d₆) δ 12.44 (s, 1H),10.53 (d, J=7.8 Hz, 1H), 8.39 (s, 1H), 7.39 (q, J=8.5 Hz, 1H), 7.32-7.14(m, 1H), 7.05 (t, J=9.1 Hz, 1H), 5.42 (dd, J=9.5, 4.2 Hz, 1H), 5.29 (p,J=6.9 Hz, 1H), 5.09 (s, 1H), 4.65 (dd, J=12.9, 4.3 Hz, 1H), 4.59 (s,1H), 4.02 (dd, J=12.6, 9.8 Hz, 1H), 1.92 (s, 4H), 1.83 (d, J=12.1 Hz,1H), 1.61-1.52 (m, 1H), 1.44 (d, J=6.9 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₂H₂₂F₂N₃O₅: 446.15; found: 446.2.

Example 14 Preparation of Compound 14(2R,5S,13aR)—N—((S)-1-(2,4-difluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 14 was prepared in a similar manner to compound 11 using(S)-1-(2,4-difluorophenyl)ethanamine in place of(R)-1-(2,4-difluorophenyl)ethanamine and using(1R,3S)-3-aminocyclopentanol in place of (1S,3R)-3-aminocyclopentanol.¹H-NMR (400 MHz, DMSO-d₆) δ 12.46 (s, 1H), 10.53 (d, J=7.6 Hz, 1H), 8.38(s, 1H), 7.39 (q, J=8.6 Hz, 1H), 7.28-7.14 (m, 1H), 7.05 (t, J=8.5 Hz,1H), 5.44 (dd, J=9.8, 3.8 Hz, 1H), 5.28 (p, J=8.0 Hz, 1H), 5.09 (s, 1H),4.66 (dd, J=12.9, 4.0 Hz, 1H), 4.59 (s, 1H), 3.99 (dd, J=12.5, 9.6 Hz,1H), 1.93 (s, 4H), 1.83 (d, J=12.6 Hz, 1H), 1.56 (dt, J=13.0, 3.3 Hz,1H), 1.45 (d, J=6.9 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₂F₂N₃O₅: 446.15; found: 446.1.

Example 15 Preparation of Compound 15(2S,5R,13aS)—N-(4-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

1-(2,2-dimethoxyethyl)-5-methoxy-6-(methoxycarbonyl)-4-oxo-1,4-dihydropyridine-3-carboxylicacid (1-A, 3.15 g, 10.0 mmol), suspended in acetonitrile (36 mL) andacetic acid (4 mL) was treated with methane sulfonic acid (0.195 mL).The mixture heated to 75° C. After 7 hours, the crude mixture was cooledand stored in a −10° C. for three days. The crude mixture was reheatedto 75° C. for 2 hours, cooled used without purification in the nextstep. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₉H₂₁F₂N₂O₇: 288.07;found: 288.1.

Step 2

Crude1-(2,2-dihydroxyethyl)-5-methoxy-6-(methoxycarbonyl)-4-oxo-1,4-dihydropyridine-3-carboxylicacid (16.8 mL of crude mixture from Step 1, approx 4 mmol) was combinedwith (1S,3R)-3-aminocyclopentanol (0.809 g, 8 mmol), diluted withacetonitrile (16.8 mL), and treated with potassium carbonate (0.553 g, 4mmol). The reaction mixture was heated to 85° C., stirred for 15minutes, cooled to ambient temperature and stirred an additional 16hours. HCl (50 mL, 0.2M aq) was added and the clear yellow solution wasextracted three times with dichloromethane. The combined organics weredried over sodium sulfate, filtered and concentrated to a yellow solid.This crude material was precipitated from dichloromethane/hexanes toafford desired intermediate 15-B as a light beige powder. ¹H-NMR (400MHz, DMSO-d₆) δ 8.72 (s, 1H), 5.42 (dd, J=9.6, 4.1 Hz, 1H), 5.09 (s,1H), 4.72 (dd, J=13.0, 3.7 Hz, 1H), 4.57 (s, 1H), 4.09 (dd, J=12.5, 9.6Hz, 1H), 3.83 (s, 3H), 1.92 (s, 3H), 1.78 (m, 2H), 1.62-1.47 (m, 1H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₅H₁₇N₂O₆: 321.11; found: 321.2.

Step 3

Intermediate 15-B (0.040 g, 0.125 mmol) and (4-fluorophenyl)methanamine(0.017 g, 0.137 mmol) were suspended in acetonitrile (1 mL) and treatedwith N,N-diisopropylethylamine (DIPEA) (0.033 mL, 0.187 mmol) and HATU(0.052 g, 0.137 mmol). After stirring for 30 minutes, the reactionmixture was treated with magnesium bromide (0.046 g, 0.25 mmol) andheated to 50° C. After 10 minutes, the reaction mixture was cooled andtreated with HCl (2 mL, 10% aq). After a few minutes, the precipitatewas filtered and washed with HCl (10/o aq) and water. Preparative HPLCpurification of the precipitate (20-65% acetonitrile:water, 0.1% TFA)afforded desired Compound 15. ¹H-NMR (400 MHz, DMSO-d₆) δ 12.44 (s, 1H),10.36 (t, J=6.0 Hz, 1H), 8.46 (s, 1H), 7.37-7.28 (m, 2H), 7.19-7.09 (m,2H), 5.43 (dd, J=9.6, 4.0 Hz, 1H), 5.08 (s, 1H), 4.68 (dd, J=12.8, 4.1Hz, 1H), 4.59 (s, 1H), 4.58-4.42 (m, 3H), 4.02 (dd, J=12.7, 9.6 Hz, 1H),1.92 (s, 5H), 1.83 (d, J=12.2 Hz, 1H), 1.56 (dt, J=12.0, 3.4 Hz, 1H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₁FN₃O₅: 414.15; found:414.2.

Example 16 Preparation of Compound 16(2S,5R,13aS)—N-(2,3-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2, I-b][1,3]oxazepine-10-carboxamide

Compound 16 was prepared in a similar manner to compound 15 using(2,3-difluorophenyl)methanamine in place of (4-fluorophenyl)methanamine.¹H-NMR (400 MHz, DMSO-d₆) δ 12.46 (s, 1H), 10.41 (t, J=6.1 Hz, 1H), 8.45(s, 1H), 7.43-7.25 (m, 1H), 7.25-7.05 (m, 2H), 5.44 (dd, J=9.5, 3.9 Hz,1H), 5.09 (s, 1H), 4.68 (dd, J=12.8, 4.0 Hz, 1H), 4.65-4.53 (m, 3H),4.02 (dd, J=12.7, 9.8 Hz, 1H), 3.56 (s, 1H), 1.93 (s, 4H), 1.83 (d,J=11.9 Hz, 1H), 1.57 (dt, J=11.5, 3.0 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₁H₂₀F₂N₃O₅: 432.14; found: 432.2.

Example 17 Preparation of Compound 17(2S,5R,13aS)—N-(4-chloro-2-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,I-b][1,3]oxazepine-10-carboxamide

Compound 17 was prepared in a similar manner to compound 15 using(4-chloro-2-fluorophenyl)methanamine in place of(4-fluorophenyl)methanamine. ¹H-NMR (400 MHz, DMSO-d₆) δ 12.46 (s, 1H),10.45-10.29 (m, 1H), 8.44 (s, 1H), 7.42 (dd, J=10.0, 2.0 Hz, 1H), 7.33(t, J=8.1 Hz, 1H), 7.26 (dd, J=8.4, 1.8 Hz, 1H), 5.50-5.38 (m, 1H), 5.09(s, 1H), 4.68 (dd, J=13.0, 4.0 Hz, 1H), 4.59 (s, 1H), 4.54 (m, 2H), 4.02(dd, J=12.8, 9.7 Hz, 1H), 1.93 (s, 4H), 1.83 (d, J=12.0 Hz, 1H), 1.57(dt, J=11.9, 3.4 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₂₀ClFN₃O₅: 448.11; found: 448.2.

Example 18 Preparation of Compound 18(2S,5R,13aS)—N-(3,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 18 was prepared in a similar manner to compound 15 using(3,4-difluorophenyl)methanamine in place of (4-fluorophenyl)methanamine.¹H-NMR (400 MHz, DMSO-d₆) δ 12.46 (s, 1H), 10.51-10.27 (m, 1H), 8.46 (s,1H), 7.50-7.23 (m, 2H), 7.23-7.03 (m, 1H), 5.44 (dd, J=9.5, 3.6 Hz, 1H),5.09 (s, 1H), 4.75-4.63 (m, 1H), 4.60 (s, 1H), 4.57-4.44 (m, 2H), 4.02(dd, J=12.6, 9.8 Hz, 1H), 1.93 (s, 4H), 1.83 (d, J=12.0 Hz, 1H), 1.57(dt, J=12.0, 3.4 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₂₀F₂N₃O₅: 432.14; found: 432.2.

Example 19 Preparation of Compound 19(1R,5S)—N-(2,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-1,5-methanopyrido[1′,2′:4,5]pyrazino[1,2-a][1,3]diazepine-10-carboxamide

Steps 1 and 2

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 97.5 mg, 0.236 mmol) was treated with acetonitrile (1.9 mL),acetic acid (0.1 mL), potassium carbonate (145 mg, 1.05 mmol), and(S)-piperidin-3-amine dihydrochloride (82 mg, 0.472 mmol). The reactionmixture was sealed and heated to 90° C. After 60 minutes, the reactionmixture was cooled partitioned between brine and dichloromethane. Theaqueous phase was thrice extracted into dichloromethane and the combinedorganic phases were combined, dried over MgSO4, filtered, concentrated.The crude product was dissolved into acetonitrile (2 mL) and magnesiumbromide (89.1 mg, 0.48 mmol) was added. The mixture was resealed andheated to 50° C. After 90 minutes, the reaction mixture was quenchedwith −5 mL of 0.2M HCl(aq), the pH adjusted to −10, diluted with brine,and thrice extracted into DCM. HPLC purification (Acetonitrile:water,0.1% TFA) afforded Compound 19. ¹H-NMR (400 MHz, Chloroform-d) δ 10.43(t, J=5.9 Hz, 1H), 8.43 (s, 1H), 7.39-7.30 (m, 1H), 6.81 (q, J=8.1 Hz,2H), 4.89 (dd, J=11.6, 3.8 Hz, 1H), 4.69 (s, 1H), 4.64 (d, J=5.8 Hz,2H), 4.26 (dd, J=12.6, 3.8 Hz, 1H), 3.91 (t, J=12.1 Hz, 1H), 3.20-3.10(m, 2H), 3.06 (s, 2H), 2.14-2.02 (m, 1H), 1.96-1.81 (m, 2H), 1.81-1.70(m, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₄O₄: 431.15;found: 431.2.

Example 20 Preparation of Compound 20(1S,5R)—N-(2,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-1,5-methanopyrido[1′,2′:4,5]pyrazino[1,2-a][1,3]diazepine-10-carboxamide

Steps 1 and 2

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 103.3 mg, 0.25 mmol) was treated with acetonitrile (1.9 mL),acetic acid (0.1 mL), potassium carbonate (159.8 mg, 1.16 mmol), and(R)-piperidin-3-amine dihydrochloride (90 mg, 0.52 mmol). The reactionmixture was sealed and heated to 90° C. After 40 minutes, the reactionmixture was cooled partitioned between brine and dichloromethane. Theaqueous phase was thrice extracted into dichloromethane and the combinedorganic phases were combined, dried over MgSO₄, filtered, concentrated.The crude product was dissolved into acetonitrile (2 mL) and magnesiumbromide (96.5 mg, 0.52 mmol) was added. The mixture was resealed andheated to 50° C. After 80 minutes, the reaction mixture was quenchedwith ˜5 mL of 0.2M HCl (aq), the pH adjusted to −10, diluted with brine,and thrice extracted into DCM. HPLC purification (Acetonitrile:water,0.1% TFA) afforded Compound 20. ¹H-NMR (400 MHz, DMSO-d₄) δ 10.35 (t,J=6.0 Hz, 1H), 8.48 (s, 1H), 7.45-7.33 (m, 1H), 7.29-7.18 (m, 1H), 7.05(td, J=8.5, 2.4 Hz, 1H), 5.06 (dd, J=11.4, 3.5 Hz, 1H), 4.56-4.47 (m,3H), 4.44 (s, 1H), 4.05 (t, J=11.8 Hz, 1H), 3.07-2.89 (m, 4H), 1.85-1.73(m, 3H), 1.54-1.46 (m, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₂₀F₂N₄O₄: 431.15; found: 431.2.

Example 21 Preparation of Compound 21(2S,5R,13aS)—N—((S)-1-(4-fluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2, I-b][1,3]oxazepine-10-carboxamide

Steps 1 and 2

(S)-Methyl1-(2,2-dihydroxyethyl)-5-(1-(4-fluorophenyl)ethylcarbamoyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(21-A, 1 mL, 0.23 M solution in 19:1 acetonitrile:acetic acid, preparedas per (R)-methyl1-(2,2-dihydroxyethyl)-5-(1-(4-fluorophenyl)ethylcarbamoyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate9-A from Example 9 using (S)-1-(4-fluorophenyl)ethanamine in place of(R)-1-(4-fluorophenyl)ethanamine) was treated with(1S,3R)-3-aminocyclopentanol (62 mg, 0.61 mmol) and potassium carbonate(34 mg, 0.25 mmol). The reaction mixture was sealed and heated to 90° C.After 60 minutes, the reaction mixture was cooled partitioned betweenbrine and dichloromethane. The aqueous phase was thrice extracted intodichloromethane and the combined organic phases were combined, driedover MgSO₄, filtered, and concentrated. The crude product was dissolvedinto acetonitrile (2 mL) and magnesium bromide (74 mg, 0.4 mmol) wasadded. The mixture was resealed and heated to 50° C. After 100 minutes,the reaction mixture was quenched with 0.2M HCl (aq), diluted withbrine, and thrice extracted into DCM. HPLC purification(acetonitrile:water, 0.1% TFA) afforded Compound 21. ¹H-NMR (400 MHz,DMSO-d₆) δ 12.42 (br s, 1H), 10.45 (d, J=7.9 Hz, 1H), 8.40 (s, 1H), 7.36(dd, J=8.6, 5.5 Hz, 2H), 7.14 (t, J=8.9 Hz, 2H), 5.42 (dd, J=9.6, 4.2Hz, 1H), 5.15-5.04 (m, 2H), 4.72-4.55 (m, 2H), 4.02 (dd, J=12.7, 9.7 Hz,1H), 1.97-1.89 (m, 4H), 1.82 (d, J=12.2 Hz, 1H), 1.56 (dt, J=11.9, 3.3Hz, 1H), 1.43 (d, J=6.9 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₂FN₃O₅: 428.16; found: 428.1.

Example 22 Preparation of Compound 22(2R,5S,13aR)—N—((S)-1-(4-fluorophenyl)ethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Steps 1 and 2

(S)-methyl1-(2,2-dihydroxyethyl)-5-(1-(4-fluorophenyl)ethylcarbamoyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(21-A, 1 mL, 0.23 M solution in 19:1 acetonitrile:acetic acid) wastreated with (1R,3S)-3-aminocyclopentanol (52 mg, 0.51 mmol) andpotassium carbonate (31 mg, 0.22 mmol). The reaction mixture was sealedand heated to 90° C. After 60 minutes, the reaction mixture was cooledpartitioned between brine and dichloromethane. The aqueous phase wasthrice extracted into dichloromethane and the combined organic phaseswere combined, dried over MgSO₄, filtered, and concentrated. The crudeproduct was dissolved into acetonitrile (2 mL) and magnesium bromide (91mg, 0.49 mmol) was added. The mixture was resealed and heated to 50° C.After 100 minutes, the reaction mixture was quenched with 0.2M HCl(aq),diluted with brine, and thrice extracted into DCM. HPLC purification(acetonitrile:water, 0.1% TFA) afforded Compound 22. ¹H-NMR (400 MHz,DMSO-d₆) δ 12.44 (br s, 1H), 10.45 (d, J=7.7 Hz, 1H), 8.39 (s, 1H), 7.36(dd, J=8.5, 5.6 Hz, 2H), 7.14 (t, J=8.9 Hz, 2H), 5.43 (dd, J=9.6, 4.0Hz, 1H), 5.15-5.06 (m, 2H), 4.66 (dd, J=12.8, 3.9 Hz, 1H), 4.58 (s, 1H),3.99 (dd, J=12.6, 9.5 Hz, 1H), 1.93 (s, 4H), 1.82 (d, J=12.0 Hz, 1H),1.56 (dt, J=12.0, 3.0 Hz, 1H), 1.44 (d, J=6.9 Hz, 3H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₂H₂₂FN₃O₅: 428.16; found: 428.1.

Example 23 Preparation of Compound 23(2S,5R,13aS)—N-(2-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Steps 1 and 2

15-B (41 mg, 0.13 mmol) was treated with acetonitrile (1 mL),(2-fluorophenyl)methanamine (17 mg, 0.14 mmol), HATU (67 mg, 0.18 mmol),and N,N-diisopropylethylamine (DIPEA) (24 mg, 0.19 mmol). The reactionmixture was stirred at room temperature for one hour and magnesiumbromide (47 mg, 0.26 mmol) was added. The mixture was sealed and heatedto 50° C. After 60 minutes, the reaction mixture was quenched with 0.2MHCl (aq), diluted with brine, and thrice extracted into DCM. HPLCpurification (Acetonitrile:water, 0.1% TFA) afforded Compound 23. ¹H-NMR(400 MHz, Chloroform-d) δ 10.42 (s, 1H), 8.34 (s, 1H), 7.36 (t, J=7.9Hz, 1H), 7.24-7.17 (m, 1H), 7.12-6.97 (m, 2H), 5.40-5.32 (m, 1H), 5.29(t, J=3.5 Hz, 1H), 4.67 (s, 3H), 4.28-4.20 (m, 1H), 4.06-3.95 (m, 1H),2.20-1.96 (m, 4H), 1.95-1.84 (m, 1H), 1.59 (dt, J=12.4, 3.3 Hz, 1H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀FN₃O₅: 414.15; found:414.2.

Example 24 Preparation of Compound 24(2S,5R,13aS)—N-(3,5-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Steps 1 and 2

15-B (44 mg, 0.14 mmol) was treated with acetonitrile (1 mL),(3,5-difluorophenyl)methanamine (32 mg, 0.23 mmol), HATU (54 mg, 0.14mmol), and N,N-diisopropylethylamine (37 mg, 0.29 mmol). The reactionmixture was stirred at room temperature for one hour and magnesiumbromide (57 mg, 0.31 mmol) was added. The mixture was sealed and heatedto 50° C. After 60 minutes, the reaction mixture was quenched with 0.2MHCl (aq), diluted with brine, and thrice extracted into DCM HPLCpurification (Acetonitrile:water, 0.1% TFA) afforded Compound 24. ¹H-NMR(400 MHz, Chloroform-d) δ 10.39 (s, 1H), 8.42 (s, 1H), 6.82 (d, J=7.9Hz, 2H), 6.65 (t, J=8.8 Hz, 1H), 5.38 (d, J=7.7 Hz, 1H), 5.28 (s, 1H),4.78-4.41 (m, 3H), 4.32 (d, J=12.1 Hz, 1H), 4.02 (t, J=10.9 Hz, 1H),2.30-1.97 (m, 4H), 1.97-1.81 (m, 1H), 1.59 (d, J=12.3 Hz, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₁₉F₂N₃O₅: 432.14; found: 432.2.

Example 25 Preparation of Compound 25(2S,5R,13aS)—N-(4-fluoro-3-(trifluoromethyl)benzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Steps 1 and 2

15-B (43 mg, 0.13 mmol) was treated with acetonitrile (1 mL),(4-fluoro-3-(trifluoromethyl)phenyl)methanamine (29 mg, 0.15 mmol), HATU(62 mg, 0.16 mmol), and N,N-diisopropylethylamine (26 mg, 0.20 mmol).The reaction mixture was stirred at room temperature for one hour andmagnesium bromide (62 mg, 0.34 mmol) was added. The mixture was sealedand heated to 50° C. After 60 minutes, the reaction mixture was quenchedwith 0.2M HCl(aq), diluted with brine, and thrice extracted into DCM.HPLC purification (Acetonitrile:water, 0.1% TFA) afforded Compound 25.¹H-NMR (400 MHz, Chloroform-d) δ 10.44 (s, 1H), 8.29 (s, 1H), 7.56-7.38(m, 2H), 7.06 (t, J=9.2 Hz, 1H), 5.30 (dd, J=9.3, 3.5 Hz, 1H), 5.21 (s,1H), 4.65-4.45 (m, 3H), 4.21 (dd, J=12.8, 3.4 Hz, 1H), 3.95 (dd, J=12.4,9.7 Hz, 1H), 2.11-1.89 (m, 4H), 1.89-1.74 (m, 1H), 1.53 (dt, J=12.4, 3.2Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₁₉F₄N₃O₅: 482.14;found: 482.2.

Example 26 Preparation of Compound 26(2S,5R,13aS)—N-(4-chloro-3-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,I-b][1,3]oxazepine-10-carboxamide

Steps 1 and 2

15-B (41 mg, 0.13 mmol) was treated with acetonitrile (1 mL),(4-chloro-3-fluorophenyl)methanamine (40 mg, 0.25 mmol), HATU (60 mg,0.16 mmol), and N,N-diisopropylethylamine (28 mg, 0.22 mmol). Thereaction mixture was stirred at room temperature for one hour andmagnesium bromide (48 mg, 0.26 mmol) was added. The mixture was sealedand heated to 50° C. After 60 minutes, the reaction mixture was quenchedwith 0.2M HCl (aq), diluted with brine, and thrice extracted into DCM.HPLC purification (Acetonitrile:water, 0.1% TFA) afforded Compound 26.¹H-NMR (400 MHz, Chloroform-d) δ 10.41 (s, 1H), 8.30 (s, 1H), 7.24 (t,J=6.1 Hz, 1H), 7.13-6.90 (m, 2H), 5.30 (dd, J=9.1, 3.2 Hz, 1H), 5.22 (s,1H), 4.61 (s, 1H), 4.51 (s, 2H), 4.20 (d, J=9.4 Hz, 1H), 3.95 (d, J=12.0Hz, 1H), 2.11-1.90 (m, 4H), 1.90-1.76 (m, 1H), 1.53 (d, J=12.2 Hz, 1H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₉ClFN₃O₅: 448.11; found:448.2.

Example 27 Preparation of Compound 27(2S,5R)—N-(1-(2,4-difluorophenyl)cyclopropyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

A suspension of the compound 1-A (1.004 g, 3.19 mmol), the amine 27-A(688 mg, 3.35 mmol), and HATU (1.453 g 3.82 mmol) in CH₂Cl₂ (20 mL) wasstirred in 0° C. bath as N,N-diisopropylethylamine (DIPEA) (2 mL, 11.48mmol) was added. After 1 hour at 0° C., the reaction mixture wasconcentrated to a syrup, diluted with ethyl acetate, and washed withwater (×2). After the aqueous fractions were extracted with ethylacetate (×1), the organic fractions were combined, dried (Na₂SO₄), andconcentrated. The residue was purified by CombiFlash (120 g column)using hexanes-ethyl acetate as eluents. The major peak was combined andconcentrated to afford 1.082 g (73%) of the product 27-B. After theminor peak was combined and concentrated, the concentrated residue wasdissolved in CH₂Cl₂ and some insoluble materials were filtered. Thefiltrate was concentrated to get 361 mg (24%) of the additional product27-B. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₂H₂₅F₂N₂O₇: 467.16;found: 467.1.

Step 2 and 3

Compound 27-B (81 mg, 0.174 mmol) was dissolved in a mixture (1 mL) ofacetonitrile (22 mL), AcOH (2 mL), and methanesulfonic acid (0.14 mL,2.16 mmol) at room temperature and the resulting solution was stirred at65° C. for 20 hours.

After the resulting solution was cooled to room temperature, theaminoalcohol 27-D (50 mg, racemic, 0.363 mmol), K₂CO₃ (50 mg, 0.362mmol), and acetonitrile (2 mL) were added to the solution. The resultingmixture was stirred at 65° C. bath for 1 hour. After the reactionmixture was cooled to room temperature, it was acidified with 1 N HCl(˜2 mL), diluted with water (˜8 mL), and extracted with CH₂Cl₂ (×3).Combined extracts were dried (Na₂SO₄), concentrated, and purified byCombiFlash to obtain 67 mg (82%) of compound 27-E. ¹H-NMR (400 MHz,CDCl₃) δ 10.53 (s, 1H), 8.25 (s, 1H), 7.60 (td, J=8.5, 6.5 Hz, 1H),6.85-6.57 (m, 2H), 5.33 (br, 1H), 5.26 (dd, J=9.6, 3.9 Hz, 1H), 4.60 (t,J=3.0 Hz, 1H), 4.18-4.06 (m, 1H), 4.01 (s, 3H), 3.92 (dd, J=12.7, 9.6Hz, 1H), 2.11-1.91 (m, 4H), 1.88-1.71 (m, 1H), 1.60-1.49 (m, 1H),1.31-1.10 (m, 4H). ¹⁹F-NMR (376.1 MHz, CDCl₃) δ −111.80 (q, J=8.8 Hz,1F), −112.05 (p, J=7.9 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₄H₂₄F₂N₃O₅: 472.17; found: 472.1.

Step 4

A mixture of compound 27-E (67 mg, 0.142 mmol) and MgBr₂ (66 mg, 0.358mmol) in MeCN (3 mL) was stirred at 50° C. for 30 minutes and cooled to0° C. before treating with 1 N HCl (3 mL). After the mixture was dilutedwith water (˜30 mL), the product was extracted with CH₂Cl₂ (×3), and thecombined extracts were dried (Na₂SO₄) and concentrated. The product waspurified by preparative HPLC and freeze-dried to obtain product 27 as a1:1 mixture with trifluoroacetic acid. ¹H-NMR (400 MHz, CDCl₃) δ 10.70(s, 1H), 8.35 (s, 1H), 7.57 (q, J=8.2 Hz, 1H), 6.91-6.56 (m, 2H), 5.31(dt, J=14.3, 4.0 Hz, 2H), 4.68 (s, 1H), 4.22 (dd, J=13.2, 3.9 Hz, 1H),3.99 (dd, J=12.8, 9.3 Hz, 1H), 2.28-1.96 (m, 5H), 1.88 (ddt, J=12.1,8.6, 3.7 Hz, 1H), 1.71-1.49 (m, 1H), 1.38-1.11 (m, 4H). ¹⁹F-NMR (376.1MHz, CDCl₃) δ −76.37 (s, 3F), −111.6˜−111.75 (m, 2F). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₃H₂₂F₂N₃O₅: 458.15; found: 458.1.

Example 28 Preparation of Compound 28(2S,6R)—N-(1-(2,4-difluorophenyl)cyclopropyl)-9-hydroxy-8,10-dioxo-3,4,5,6,8,10,14,14a-octahydro-2H-2,6-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazocine-11-carboxamide

Step 1 and 2

Compound 27-B (87 mg, 0.187 mmol) was dissolved in a mixture (2 mL) ofacetonitrile (22 mL), AcOH (2 mL), and methanesulfonic acid (0.14 mL,2.16 mmol) at room temperature and the resulting solution was stirred at65° C. for 20 hours.

After the resulting solution was cooled to room temperature, theaminoalcohol 28-A (44 mg, racemic, 0.382 mmol) and acetonitrile (2 mL)were added to the solution. After the resulting mixture was stirred at65° C. bath for 30 minutes, K₂CO₃ (41 mg, 0.297 mmol) was added and themixture was stirred at 65° C. for 21 hours. The reaction mixture wascooled to room temperature, it was acidified with 1 N HCl (˜2 mL),diluted with water (˜8 mL), and extracted with CH₂Cl₂ (×3). Combinedextracts were dried (Na₂SO₄), concentrated, and purified by preparativeHPLC and the fraction containing the product was freeze-dried. After theresidue was dissolved in ethyl acetate, the solution was washed withsaturated NaHCO₃ (×1), dried (Na₂SO₄), and concentrated to obtain 18 mg(20%) of compound 28-B as a 1:1 mixture with trifluoroacetic acid.¹H-NMR (400 MHz, CDCl₃) δ 10.54 (s, 1H), 8.26 (s, 1H), 7.63 (td, J=8.6,6.6 Hz, 1H), 6.76 (dddd, J=21.9, 11.2, 8.7, 2.3 Hz, 2H), 5.39 (dd,J=9.6, 3.7 Hz, 1H), 4.53-4.36 (m, 2H), 4.09 (dd, J=12.8, 3.7 Hz, 1H),4.03 (s, 3H), 3.99 (dd, J=12.7, 9.7 Hz, 1H), 2.41-2.20 (m, 2H), 1.84(dtd, J=19.7, 9.3, 8.8, 4.4 Hz, 2H), 1.74 (dd, J=14.6, 2.5 Hz, 1H),1.62-1.35 (m, 2H), 1.34-1.14 (m, 5H). ¹⁹F-NMR (376.1 MHz, CDCl₃) δ−111.75 (q, J=8.9 Hz, 1F), −112.01 (p, J=7.9 Hz, 1F). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₅H₂₆F₂N₃O₅: 486.18; found: 486.2.

Step 3

Compound 28-B (18 mg, 0.037 mmol) was treated with MgBr₂ as described instep 4 in the synthesis of compound 27-E to obtain compound 28. ¹H-NMR(400 MHz, CDCl₃) δ 10.66 (s, 1H), 8.29 (s, 1H), 7.59 (td, J=8.5, 6.6 Hz,1H), 6.89-6.60 (m, 2H), 5.51 (dd, J=9.9, 4.0 Hz, 1H), 4.55 (s, 1H), 4.48(t, J=4.2 Hz, 1H), 4.21 (dd, J=12.9, 4.1 Hz, 1H), 3.99 (dd, J=12.8, 9.8Hz, 1H), 2.56-2.35 (m, 1H), 2.14 (dd, J=16.1, 5.9 Hz, 1H), 1.96-1.74 (m,3H), 1.66-1.37 (m, 3H), 1.28 (d, J=4.4 Hz, 2H), 1.26-1.19 (m, 2H).¹⁹F-NMR (376.1 MHz, CDCl₃) δ −76.41 (s, 3F, −111.79 (m, 2F). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₄H₂₃F₂N₃O₅: 472.17; found: 472.1.

Example 29 Preparation of Compound 29(2R,6S)—N-(1-(2,4-difluorophenyl)cyclopropyl)-9-hydroxy-8,10-dioxo-3,4,5,6,8,10,14,14a-octahydro-2H-2,6-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazocine-11-carboxamide

Step 1 and 2

Compound 29-B (13 mg, 14%) was prepared from compound 27-B (87 mg, 0.187mmol) and the aminoalcohol 29-A (45 mg, 0.391 mmol) in a manner similarto that described in step 1 of the synthesis of compound 28-B. ¹H-NMR(400 MHz, CDCl₃) δ 10.54 (s, 1H), 8.26 (s, 1H), 7.63 (td, J=8.6, 6.6 Hz,1H), 6.76 (dddd, J=21.9, 11.2, 8.7, 2.3 Hz, 2H), 5.39 (dd, J=9.6, 3.7Hz, 1H), 4.53-4.36 (m, 2H), 4.09 (dd, J=12.8, 3.7 Hz, 1H), 4.03 (s, 3H),3.99 (dd, J=12.7, 9.7 Hz, 1H), 2.41-2.20 (m, 2H), 1.84 (dtd, J=19.7,9.3, 8.8, 4.4 Hz, 2H), 1.74 (dd, J=14.6, 2.5 Hz, 1H), 1.62-1.35 (m, 2H),1.34-1.14 (m, 5H). ¹⁹F-NMR (376.1 MHz, CDCl₃) δ −111.75 (q, J=8.9 Hz,1F), −112.01 (p, J=7.9 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₅H₂₆F₂N₃O₅: 486.18; found: 486.2.

Step 3

Compound 29 was prepared from compound 29-B in a manner similar to thatdescribed in step 2 of the synthesis of compound 16. ¹H-NMR (400 MHz,CDCl₃) δ 10.66 (s, 1H), 8.29 (s, 1H), 7.59 (td, J=8.5, 6.6 Hz, 1H),6.89-6.60 (m, 2H), 5.51 (dd, J=9.9, 4.0 Hz, 1H), 4.55 (s, 1H), 4.48 (t,J=4.2 Hz, 1H), 4.21 (dd, J=12.9, 4.1 Hz, 1H), 3.99 (dd, J=12.8, 9.8 Hz,1H), 2.56-2.35 (m, 1H), 2.14 (dd, J=16.1, 5.9 Hz, 1H), 1.96-1.74 (m,3H), 1.66-1.37 (m, 3H), 1.28 (d, J=4.4 Hz, 2H), 1.26-1.19 (m, 2H).¹⁹F-NMR (376.1 MHz, CDCl₃) δ −76.41 (s, 3F, −111.79 (m, 2F). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₄H₂₃F₂N₃O₅: 472.17; found: 472.1.

Example 30 Preparation of Compound 30(2S,5R,13aS)—N-(1-(2,4-difluorophenyl)cyclopropyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1 and 2

Compound 27-B (150 mg, 0.322 mmol) was dissolved in acetonitrile (2 mL),AcOH (0.2 mL), and methanesulfonic acid (0.007 mL, 0.108 mmol) at roomtemperature and the resulting solution was stirred at 65° C. for 20hours. After the resulting solution was cooled to room temperature, theaminoalcohol 30-A (72.1 mg, chiral, 0.713 mmol), K₂CO₃ (89.4 mg, 0.647mmol), and acetonitrile (2 mL) were added to the solution. The resultingmixture was stirred at 65° C. bath for 0.5 hour. After the reactionmixture was cooled to room temperature, it was acidified with 1 N HCl(˜3 mL), diluted with water (˜12 mL), and extracted with CH₂Cl₂ (×3).Combined extracts were dried (Na₂SO₄), concentrated, and purified byCombiFlash to obtain 128 mg (84%) of compound 30-B. ¹H-NMR (400 MHz,CDCl₃) δ 10.52 (s, 1H), 8.24 (s, 1H), 7.61 (td, J=8.6, 6.6 Hz, 1H),6.85-6.65 (m, 2H), 5.33 (t, J=4.1 Hz, 1H), 5.25 (dd, J=9.5, 3.9 Hz, 1H),4.61 (d, J=3.4 Hz, 1H), 4.18-4.08 (m, 1H), 4.02 (s, 3H), 3.99-3.87 (m,1H), 2.12-1.91 (m, 4H), 1.85-1.69 (m, 1H), 1.55 (ddd, J=12.3, 4.1, 2.8Hz, 1H), 1.31-1.14 (m, 4H). ¹⁹F-NMR (376.1 MHz, CDCl₃) δ −111.79 (q,J=8.8 Hz, 1F), −112.05 (p, J=7.9 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₄H₂₄F₂N₃O₅: 472.17; found: 472.2.

Step 3

A mixture of compound 30-B (128 mg, 0.272 mmol) and MgBr₂ (130 mg, 0.706mmol) in MeCN (5 mL) was stirred at 50° C. for 30 minutes and cooled to0° C. before treating with 1 N HCl (4 mL). After the mixture was dilutedwith water, the product was extracted with CH₂Cl₂ (×3), and the combinedextracts were dried (Na₂SO₄) and concentrated. The product was purifiedby CombiFlash to obtain product 30. ¹H-NMR (400 MHz, CDCl₃) δ 12.27 (s,1H), 10.52 (s, 1H), 8.16 (s, 1H), 7.61 (td, J=8.6, 6.6 Hz, 1H),6.96-6.54 (m, 2H), 5.36-5.23 (m, 2H), 4.66 (t, J=3.1 Hz, 1H), 4.18-4.06(m, 1H), 3.94 (dd, J=12.8, 9.4 Hz, 1H), 2.20-1.95 (m, 4H), 1.89 (td,J=11.4, 9.8, 6.7 Hz, 1H), 1.70-1.54 (m, 1H), 1.32-1.15 (m, 4H). ¹⁹F-NMR(376.1 MHz, CDCl₃) δ −111.87 (q, J=8.9 Hz, 1F), −112.21 (p, J=7.9 Hz,1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₃H₂₂F₂N₃O₅: 458.15; found:458.2.

Example 31 Preparation of Compound 31(2R,5S)—N-(1-(2,4-difluorophenyl)cyclopropyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1 and 2

Compound 31-B (123 mg, 81%) was prepared from compound 27-B (150 mg,0.322 mmol) and the aminoalcohol 31-A (70.3 mg, 0.695 mmol) in a mannersimilar to that described in step 1 and 2 of the synthesis of compound30-B. ¹H-NMR (400 MHz, CDCl₃) δ 10.52 (s, 1H), 8.24 (s, 1H), 7.62 (td,J=8.6, 6.6 Hz, 1H), 6.91-6.63 (m, 2H), 5.33 (t, J=4.1 Hz, 1H), 5.25 (dd,J=9.5, 3.9 Hz, 1H), 4.61 (d, J=3.4 Hz, 1H), 4.14-4.07 (m, 1H), 4.03 (s,3H), 3.93 (dd, J=12.7, 9.5 Hz, 1H), 2.12-1.91 (m, 4H), 1.85-1.69 (m,1H), 1.55 (ddd, J=12.3, 4.1, 2.8 Hz, 1H), 1.31-1.14 (m, 4H). ¹⁹F-NMR(376.1 MHz, CDCl₃) δ −111.79 (q, J=9.2, 8.7 Hz, 1F), −112.03 (h, J=8.1,7.5 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₄H₂₄F₂N₃O: 472.17;found: 472.1.

Step 3

Compound 31 was prepared from compound 31-B in a manner similar to thatdescribed in step 3 of the synthesis of compound 30. ¹H-NMR (400 MHz,CDCl₃) δ 12.26 (s, 1H), 10.49 (s, 1H), 8.13 (s, 1H), 7.58 (td, J=8.6,6.5 Hz, 1H), 6.90-6.56 (m, 2H), 5.32 (dd, J=9.4, 4.1 Hz, 1H), 5.27-5.22(m, 1H), 4.64 (t, J=3.1 Hz, 1H), 4.11 (dd, J=12.8, 4.0 Hz, 1H),4.01-3.79 (m, 1H), 2.28-1.95 (m, 4H), 1.95-1.80 (m, 1H), 1.71 (m, 1H),1.56 (m, 1H), 1.42-1.08 (m, 4H). ¹⁹F-NMR (376.1 MHz, CDCl₃) δ −111.95(q, J=8.9 Hz, 1F), −112.22 (p, J=7.9 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₃H₂₂F₂N₃O₅: 458.15; found: 458.1.

Example 32 Preparation of Compound 32(2S,5R)—N-(1-(2,4-difluorophenyl)cyclobutyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

A solution of compound 32-A (22.2 mg, 0.069 mmol), compound 32-B (18.7mg, 0.102 mmol), and HATU (43 mg, 0.113 mmol) in CH₂Cl₂ (2 mL) wasstirred at room temperature as N,N-diisopropylethylamine (DIPEA) (0.075mL, 0.431 mmol) was added. After 30 minutes, the reaction mixture wasdiluted with ethyl acetate and washed with water (×2). After the aqueousfractions were extracted with EA (×1), the organic fractions werecombined, dried, concentrated, and dried in vacuum.

A mixture of the above crude product and MgBr₂ (35 mg, 0.190 mmol) inMeCN (2 mL) was stirred at 50° C. bath for 1 hour and cooled to 0° C.before being treated with 1 N HCl (˜1 mL). The resulting solution wasdiluted with water, and extracted with CH₂Cl₂ (×3). The combinedextracts were dried (Na₂SO₄), and concentrated. The product was purifiedby preparative HPLC and freeze-dried to obtain compound 32. ¹H-NMR (400MHz, CDCl₃) δ 10.87 (s, 1H), −9.3 (br, 1H), 8.35 (s, 1H), 7.50 (td,J=8.7, 6.3 Hz, 1H), 6.89-6.78 (m, 1H), 6.72 (ddd, J=11.2, 8.9, 2.6 Hz,1H), 5.48-5.12 (m, 2H), 4.72-4.60 (m, 1H), 4.22 (dd, J=13.0, 4.1 Hz,1H), 3.98 (dd, J=12.9, 9.4 Hz, 1H), 2.68 (m, 4H), 2.33-1.98 (m, 6H),1.90 (m, 2H), 1.60 (ddd, J=12.4, 4.1, 2.7 Hz, 1H). ¹⁹F-NMR (376.1 MHz,CD₃CN) δ −76.39 (s, 3F), −110.50 (q, J=9.2 Hz, 1F), −112.65 (p, J=7.8Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₄H₂₄F₂N₃O₅: 472.17;found: 472.0.

Example 33 Preparation of Compound 33(2S,5R)—N-(1-(2,4-difluorophenyl)cyclopentyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 33 was obtained from compound 32-A and compound 33-A asdescribed in the synthesis of compound 32. ¹H-NMR (400 MHz, CDCl₃) δ10.70 (s, 1H), −9.5 (br, 1H), 8.41 (s, 1H), 7.43 (td, J=8.9, 6.4 Hz,1H), 6.85-6.76 (m, 1H), 6.72 (ddd, J=11.5, 8.8, 2.6 Hz, 1H), 5.48-5.18(m, 2H), 4.68 (t, J=3.2 Hz, 1H), 4.26 (dd, J=13.0, 4.1 Hz, 1H), 4.00(dd, J=13.0, 9.4 Hz, 1H), 2.72-2.45 (m, 2H), 2.22-1.96 (m, 6H),1.96-1.75 (m, 5H), 1.60 (ddd, J=12.5, 4.1, 2.7 Hz, 1H). ¹⁹F-NMR (376.1MHz, CD₃CN) δ −76.41 (s, 3F), −107.86 (q, J=9.4 Hz, 1F), −113.13 (p,J=8.0 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₅H₂₆F₂N₃O₅:486.18; found: 485.9.

Example 34 Preparation of Compound 34(2S,5R)—N-(1-(2,4-difluorophenyl)cyclohexyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 34 was obtained from compound 32-A and compound 34-A asdescribed in the synthesis of compound 32. ¹H-NMR (400 MHz, CDCl₃) δ10.83 (s, 1H), −9.6 (br, 1H), 8.44 (s, 1H), 7.37 (td, J=9.0, 6.4 Hz,1H), 6.97-6.76 (m, 1H), 6.69 (ddd, J=11.9, 8.8, 2.7 Hz, 1H), 5.48-5.18(m, 2H), 4.68 (t, J=3.0 Hz, 1H), 4.28 (dd, J=13.1, 4.1 Hz, 1H), 4.03(dd, J=13.0, 9.4 Hz, 1H), 2.60 (d, J=13.1 Hz, 2H), 2.29-1.96 (m, 4H),1.95-1.77 (m, 4H), 1.77-1.65 (m, 4H), 1.61 (ddd, J=12.5, 4.1, 2.7 Hz,1H), 1.30 (br, 1H). ¹⁹F-NMR (376.1 MHz, CD₃CN) δ −76.41 (s, 3F), −107.86(q, J=9.4 Hz, 1F), −113.13 (p, J=8.0 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₆H₂₈F₂N₃O₅: 500.20; found: 500.0.

Example 35 Preparation of Compound 35(2S,5R)—N-(4-(2,4-difluorophenyl)tetrahydro-2H-pyran-4-yl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 35 was obtained from compound 32-A and compound 35-A asdescribed in the synthesis of compound 32. ¹H-NMR (400 MHz, CDCl₃) δ10.95 (s, 1H), 8.33 (s, 1H), −7.6 (br, 1H), 7.38 (td, J=9.0, 6.3 Hz,1H), 6.85 (td, J=8.4, 2.6 Hz, 1H), 6.73 (ddd, J=11.7, 8.6, 2.6 Hz, 1H),5.32 (dt, J=14.4, 4.0 Hz, 2H), 4.68 (t, J=3.1 Hz, 1H), 4.24 (dd, J=13.0,3.9 Hz, 1H), 4.11-3.81 (m, 5H), 2.60 (d, J=13.7 Hz, 2H), 2.33-2.17 (m,2H), 2.18-1.97 (m, 4H), 1.87 (m, 1H), 1.61 (dt, J=12.5, 3.3 Hz, 1H).¹⁹F-NMR (376.1 MHz, CD₃CN) δ −76.40 (s, 3F), −108.78 (q, J=10.3, 9.8 Hz,1F), −112.63 (p, J=8.0 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₅H₂₆F₂N₃O₆: 502.18; found: 502.0.

Example 36 Preparation of Compound 36(2S,5R)—N—((S)-1-(2,4-difluorophenyl)-2,2,2-trifluoroethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 36 was obtained from compound 32-A and compound 36-A asdescribed in the synthesis of compound 32. ¹H-NMR (400 MHz, CDCl₃) δ11.31 (d, J=9.4 Hz, 1H), 8.41 (s, 1H), 7.65-7.44 (m, 1H), 6.95 (ddd,J=9.6, 5.6, 2.0 Hz, 1H), 6.92-6.79 (m, 1H), 6.15 (h, J=7.4 Hz, 1H), −6(br, 1H), 5.41 (dd, J=9.5, 4.0 Hz, 1H), 5.31 (t, J=4.0 Hz, 1H), 4.70 (s,1H), 4.34 (dd, J=12.8, 3.9 Hz, 1H), 4.05 (dd, J=12.9, 9.4 Hz, 1H),2.26-1.99 (m, 4H), 1.99-1.87 (m, 1H), 1.62 (dt, J=12.6, 3.4 Hz, 1H).¹⁹F-NMR (376.1 MHz, CDCl₃) δ −75.23 (t, J=6.9 Hz, 3F), −76.33 (s, 3F),−108.31 (m, 1F), −112.30 (p, J=8.0 Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₂H₁₉F₅N₃O₅: 500.12; found: 500.1.

Example 37 Preparation of Compound 37(3S,11aR)—N-(1-(2,4-difluorophenyl)cyclopropyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydrooxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide

Step 1

Methyl5-(1-(2,4-difluorophenyl)cyclopropylcarbamoyl)-1-(2,2-dimethoxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(27-B, 0.150 g, 0.32 mmol) in acetonitrile (1.5 mL) and acetic acid (0.2mL) was treated with methanesulfonic acid (0.05 mL), sealed with ayellow cap, and heated to 70° C. After 16 hours, the mixture was cooledto afford a crude solution of methyl5-(1-(2,4-difluorophenyl)cyclopropylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate27-C. LCMS-ESI⁺ (m): [M+H]⁺ calculated for C₁₈H₁₉F₂N₂O₇: 439; found:439.

Steps 2 and 3

Methyl5-(1-(2,4-difluorophenyl)cyclopropylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(27-C, 0.32 mmol, the crude mixture from the previous step) wasdissolved in acetonitrile (1.5 mL) and acetic acid (0.2 mL).(S)-2-aminopropan-1-ol (0.048 g, 0.64 mmol) and K₂CO₃ (0.088 g, 0.64mmol) were added to the reaction mixture. The reaction mixture wassealed and heated to 70° C. After 3 hours, the reaction mixture wascooled and magnesium bromide (0.081 g, 0.44 mmol) was added. The mixturewas resealed and heated to 50° C. After 10 minutes, the reaction mixturewas cooled to 0° C. and 1 N hydrochloric acid (0.5 mL) was added in.Then the reaction mixture was diluted with MeOH (2 mL). Afterfiltration, the crude was purified by Prep-HPLC (30-70%acetonitrile:water, 0.1% TFA) to afford Compound 37 as a TFA salt.¹H-NMR (400 MHz, Methanol-d₄) δ 8.31 (s, 1H), 7.62 (td, J=9.2, 8.7, 6.5Hz, 1H), 7.02-6.78 (m, 2H), 5.53-5.20 (m, 1H), 4.68 (dd, J=12.3, 4.2 Hz,1H), 4.40 (dq, J=19.1, 6.7 Hz, 2H), 3.98 (dd, J=12.2, 10.0 Hz, 1H), 3.71(dd, J=8.3, 6.3 Hz, 1H), 1.41 (d, J=6.1 Hz, 3H), 1.22 (s, 4H). ¹⁹F-NMR(376 MHz, Methanol-d₄) δ −113.66˜−113.95 (m, 1F), −113.94˜−114.29 (m,1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₅: 432; found:432.

Example 38 Preparation of Compound 38(1S,4R,12aR)—N-(2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A solution of compound 38-A (1562 mg, 5.799 mmol) (see Example 41b in WO97/05139) in THF (10 mL) was stirred at −78° C. as 2.0 M LiBH₄ in THF(3.2 mL) was added and the resulting mixture was stirred at roomtemperature. After 3 hours, additional 2.0 M LiBH₄ in THF (3.2 mL) wasadded and the solution was stirred at room temperature for 17.5 hours.After the reaction mixture was diluted with ethyl acetate and addedwater slowly, two phases were separated, and the separated aqueousfraction was extracted with ethyl acetate (×1). Two organic fractionswere washed with water (×1), combined, dried (Na₂SO₄), and concentrated.The residue was purified by CombiFlash (40 g column) using hexanes-ethylacetate as eluents to afford compound 38-B. ¹H-NMR (400 MHz,Chloroform-d) δ 4.11 (s, 1H), 3.65-3.52 (m, 2H), 3.45 (m, 1H), 2.32 (d,J=4.1 Hz, 1H), 2.20 (s, 1H), 1.75-1.64 (m, 2H), 1.61 (m, 2H), 1.49-1.41(m, 1H), 1.47 (s, 9H), 1.28-1.23 (d, J=10 Hz, 1H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₁₂H₂₂NO₃: 228.16; found: 227.7.

Step 2

A solution of compound 38-B (589 mg, 2.591 mmol) and NEt₃ (0.47 mL,3.369 mmol) in CH₂Cl₂ (6 mL) was stirred at 0° C. as MsCl (0.22 mL,2.842 mmol) was added. After 1 hour at room temperature, the mixture wasdiluted with ethyl acetate and washed with water (×2). The aqueousfractions were extracted with ethyl acetate (×1), and the organicfractions were combined, dried (Na₂SO₄), and concentrated. The residuewas purified by Combi Flash (40 g column) using hexanes-ethyl acetate aseluents to afford compound 38-C. ¹H-NMR (400 MHz, Chloroform-d) δ4.39-4.28 (m, 1H), 4.16 (s, 0.4H), 4.06 (s, 0.6H), 3.98 (dd, J=10.0, 8.7Hz, 0.6H), 3.86 (t, J=9.6 Hz, 0.4H), 3.51 (dd, J=9.3, 3.7 Hz, 0.6H),3.43 (dd, J=9.3, 3.6 Hz, 0.4H), 3.02 (s, 3H), 2.59 (m, 1H), 1.82-1.58(m, 4H), 1.51-1.44 (m, 9H), 1.41 (d, J=14.8 Hz, 1H), 1.31 (s, 0.6H),1.29 (s, 0.4H).

Step 3

To a solution of compound 38-C (769 mg, 2.518 mmol,) in DMF (5 mL) wasadded sodium azide (819 mg, 12.6 mmol). The reaction mixture was stirredat 50° C. for 15 hours, at 80° C. for 5 hours, and at 100° C. for 19hours. The reaction mixture was diluted with 5% LiCl solution and theproduct was extracted with ethyl acetate (×2). After the organicfractions were washed with water (×1), the two organic fractions werecombined, dried (Na₂SO₄), and concentrated. The residue was purified byCombiFlash (40 g column) using hexanes-ethyl acetate as eluents toafford compound 38-D. ¹H-NMR (400 MHz, Chloroform-d) δ 4.16 (s, 0.4H),4.06 (s, 0.6H), 3.61 (dd, J=12.2, 3.6 Hz, 0.6H), 3.51 (dd, J=12.1, 3.2Hz, 0.4H), 3.38 (dd, J=9.4, 3.4 Hz, 0.6H), 3.26 (dd, J=9.8, 3.3 Hz,0.4H), 3.06 (dd, J=12.2, 9.4 Hz, 0.6H), 3.01-2.92 (m, 0.4H), 2.48 (d,J=5.2 Hz, 1H), 1.82-1.57 (m, 4H), 1.46 (d, J=3.0 Hz, 9H), 1.42 (m, 1H),1.28 (m, 0.6H), 1.27-1.23 (m, 0.4H).

Step 4

To a solution of compound 38-D (507 mg, 2.009 mmol,) in ethyl acetate(10 mL) and EtOH (10 mL) was added 10% Pd/C (52 mg). The reactionmixture was stirred under H₂ atmosphere for 1.5 hours. The mixture wasfiltered through celite and the filtrate was concentrated to affordcrude compound 38-E. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₂H₂₃N₂O₂:227.18; found: 226.8.

Step 5

The mixture of crude compound 38-E (206 mg, 0.910 mmol), compound 38-F(330 mg, 0.953 mmol), and NaHCO₃ (154 mg, 1.833 mmol) in water (3 mL)and EtOH (3 mL) was stirred at room temperature for 20 hours. After thereaction mixture was diluted with water and extracted with ethyl acetate(×2), the extracts were washed with water (×1), combined, dried(Na₂SO₄), and concentrated to afford the crude pyridine product.

The crude residue (388 mg) was dissolved in CH₂Cl₂ (4 mL) and 4 N HCl indioxane (4 mL). After 1.5 hours, additional 4 N HCl in dioxane (4 mL)was added and stirred for 1 hour at room temperature. The mixture wasconcentrated to dryness, coevaporated with toluene (×1) and dried invacuum for 30 minutes.

The crude residue and 1,8-diazabicycloundec-7-ene (DBU) (1.06 mL, 7.088mmol) in toluene (10 mL) was stirred at 110° C. bath. After 30 minutes,the mixture was concentrated and the residue was purified by CombiFlash(40 g column) using ethyl acetate-20% MeOH/ethyl acetate as eluents toobtain compound 38-G. ¹H-NMR (400 MHz, Chloroform-d) δ 8.03 (s, 1H),7.68-7.58 (m, 2H), 7.36-7.27 (m, 3H), 5.53 (d, J=9.9 Hz, 1H), 5.11 (d,J=9.9 Hz, 1H), 4.93 (s, 1H), 4.43-4.30 (m, 2H), 3.89 (dd, J=12.2, 3.3Hz, 1H), 3.73 (t, J=12.0 Hz, 1H), 3.59 (dd, J=11.9, 3.3 Hz, 1H), 2.53(d, J=2.8 Hz, 1H), 1.87-1.67 (m, 4H), 1.55 (d, J=10.0 Hz, 1H), 1.51-1.45(m, 1H), 1.38 (t, J=7.1 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₃H₂₅N₂O₅: 409.18; found: 409.2.

Step 6

The mixture of compound 38-G (232 mg, 0.568 mmol) in THF (3 mL) and MeOH(3 mL) was stirred at room temperature as 1 N KOH (3 mL) was added.After 1 hour, the reaction mixture was neutralized with 1 N HCl (˜3.1mL), concentrated, and the residue was concentrated with toluene (×3).After the residue was dried in vacuum for 30 minutes, a suspension ofthe crude residue, 2,4-difluorobenzylamine (86 mg, 0.601 mmol), and HATU(266 mg, 0.700 mmol) were in CH₂Cl₂ (4 mL) and DMF (4 mL) was stirred at0° C. as N,N-diisopropylethylamine (DIPEA) (0.7 mL, 4.019 mmol) wasadded. After 45 minutes, additional 2,4-difluorobenzylamine (86 mg,0.559 mmol), HATU (266 mg, 0.700 mmol), and N,N-diisopropylethylamine(DIPEA) (0.7 mL, 4.019 mmol) were added at room temperature. After 1.25hours, the mixture was concentrated to remove most of CH₂Cl₂, dilutedwith ethyl acetate, and washed with 5% LiCl (×2). After the aqueousfractions were extracted with ethyl acetate (×1), the organic fractionswere combined, dried (Na2SO4), and concentrated. The residue waspurified by Combiflash (40 g column) using ethyl acetate −20% MeOH/ethylacetate as eluents to afford compound 38-H. ¹H-NMR (400 MHz,Chloroform-d) δ 10.48 (t, J=6.0 Hz, 1H), 8.33 (s, 1H), 7.62-7.51 (m,2H), 7.40-7.27 (m, 4H), 6.87-6.75 (m, 2H), 5.39 (d, J=10.0 Hz, 1H), 5.15(d, J=10.0 Hz, 1H), 4.92 (s, 1H), 4.68-4.53 (m, 2H), 3.97 (dd, J=12.5,3.4 Hz, 1H), 3.77 (t, J=12.2 Hz, 1H), 3.55 (dd, J=12.1, 3.3 Hz, 1H),2.53 (d, J=3.1 Hz, 1H), 1.88-1.62 (m, 4H), 1.59-1.42 (m, 2H). ¹⁹F-NMR(376 MHz, Chloroform-d) δ −112.17 (q, J=7.6 Hz, 1F), −114.79 (q, J=8.6Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₈H26F2N3O4: 506.19;found: 506.2.

Step 7

Compound 38-H (240 mg, 0.475 mmol) was dissolved in TFA (3 mL) at roomtemperature for 30 minutes, and the solution was concentrated. Theresidue was purified by CombiFlash (40 g column) using CH₂Cl₂-20%0 MeOHin CH₂Cl₂ as eluents. After the collected product fractions wereconcentrated, the residue was triturated in MeCN (˜2 mL) at 0° C. for 15minutes, and the solids were filtered and washed with MeCN. Thecollected solids were dried in vacuum to afford compound 38.

The filtrate was concentrated, and the residue was dissolved in MeCN (˜1mL) and water (˜1 mL) by heating. The solution was slowly cooled to roomtemperature and then in ice bath for 15 minutes. The solids werefiltered and washed with MeCN, and dried in vacuum to afford additionalcompound 38. ¹H-NMR (400 MHz, Chloroform-d) δ 11.68 (s, 1H), 10.42 (s,1H), 8.27 (s, 1H), 7.41-7.31 (m, 1H), 6.86-6.73 (m, 2H), 4.90 (d, J=2.5Hz, 1H), 4.71-4.53 (m, 2H), 4.07 (d, J=10.6 Hz, 1H), 3.90-3.67 (m, 2H),2.68 (s, 1H), 2.01 (s, 1H), 1.97-1.80 (m, 3H), 1.80-1.62 (m, 2H).¹⁹F-NMR (376 MHz, Chloroform-d) δ −112.28 (m, 1F), −114.74 (m, 1F).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₉F₂N₃O₄: 416.14; found:416.3.

Examples 39 and 40 Preparation of Compounds 39 and 40(2R,3S,5R,13aS)—N-(2,4-difluorobenzyl)-8-hydroxy-3-methyl-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide39 and(2S,3R,5S,13aR)—N-(2,4-difluorobenzyl)-8-hydroxy-3-methyl-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide40

Step 1

Cuprous cyanide (290 mg, 3.27 mmol) was suspended in 3.3 mL THF andcooled to −78° C. A 1.6M solution of MeLi (4.1 mL, 6.56 mmol) in diethylether was added dropwise, the reaction solution was allowed to warm toroom temperature over the course of 2 hours, and recooled to −78° C.Tert-butyl (1R,3R,5S)-6-oxabicyclo[3.1.0]hexan-3-ylcarbamate (330 mg,1.66 mmol) was added dropwise in 3.3 mL THF, followed by borontrifluoride diethyl etherate (0.25 mL, 1.99 mmol), allowed to warm to−30° C. over 30 minutes, and stirred between −35° C. and −25° C. for onehour. The reaction solution was then warmed to room temperature andquenched with a mixture of saturated NH₃(aq)/NH₄(aq), extracted toEtOAc, washed with brine, dried over MgSO4, filtered, concentrated, andpurified by SGC (0-10% EtOH/DCM) to afford racemic tert-butyl(1S,3S,4S)-3-hydroxy-4-methylcyclopentylcarbamate. ¹H-NMR (400 MHz,Chloroform-d) δ 5.16 (s, 1H), 3.98 (s, 1H), 3.74 (q, J=4.3 Hz, 1H), 3.65(q, J=7.0 Hz, 1H), 2.23 (dt, J=14.0, 7.0 Hz, 1H), 1.98 (dt, J=13.3, 7.0Hz, 1H), 1.89-1.79 (m, 1H), 1.58-1.44 (m, 1H), 1.38 (s, 9H), 1.18 (t,J=7.0 Hz, 1H), 0.91 (d, J=7.0 Hz, 3H).

Step 2

3 mL HCl/dioxane (4M, 12 mmol) was added to a solution of racemictert-butyl (1S,3S,4S)-3-hydroxy-4-methylcyclopentylcarbamate (182 mg,0.85 mmol) in 3 mL dioxane. The reaction mixture was stirred at roomtemperature for 2 hours, concentrated and twice chased with toluene toafford racemic (1S,2S,4S)-4-amino-2-methylcyclopentanol.

Step 3

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 310 mg, 0.75 mmol), racemic(1S,2S,4S)-4-amino-2-methylcyclopentanol (115 mg, 0.76 mmol), andpotassium carbonate (232 mg, 1.68 mmol) were taken up in 3.8 mLacetonitrile/0.2 mL acetic acid and stirred at 90° C. for 2 hours, afterwhich the reaction mixture was partitioned between DCM and brine, theaqueous phase extracted to DCM, combined organic phases dried overMgSO₄, filtered, concentrated, and purified by SGC (0-10% EtOH/DCM) toafford intermediate 39-A.

Step 4

Intermediate 39-A (190 mg) was separated by chiral Prep-HPLC on a LuxCellulose-2 column using 9:1 ACN:MeOH as eluent to afford Intermediates39-B (first eluting peak) and 40-A (second eluting peak) inenantioenriched form. For intermediate 39-B: (absolute stereochemistryconfirmed by XRay crystallography), Chiral HPLC retention time=3.98minutes (Lux Cellulose-2 IC, 150×4.6 mm, 2 mL/min 9:1 ACN:MeOH). Forintermediate 40-A: (absolute stereochemistry confirmed by XRaycrystallography), Chiral HPLC retention time=6.35 minutes (LuxCellulose-2 IC, 150×4.6 mm, 2 mL/min 9:1 ACN:MeOH).

Step 5a

Magnesium bromide (68 mg, 0.37 mmol) was added to a solution ofintermediate 39-B (83 mg, 0.18 mmol) in 2 mL acetonitrile. The reactionmixture was stirred at 50° C. for 1 hour, acidified with 10% aqueousHCl, partitioned between the aqueous and dichloromethane, and theaqueous phase extracted to dichloromethane. The combined organic phaseswere dried over MgSO4, filtered, concentrated, and purified by silicagel chromatography (0-10% EtOH/DCM) to afford compound 39. ¹H-NMR (400MHz, Chloroform-d) δ 12.32 (s, 1H), 10.36 (s, 1H), 8.29 (s, 1H),7.44-7.33 (m, 1H), 6.88-6.76 (m, 2H), 5.37 (dd, J=9.5, 4.1 Hz, 1H), 5.28(t, J=5.3 Hz, 1H), 4.63 (d, J=5.9 Hz, 2H), 4.23 (d, J=23.0 Hz, 2H), 3.99(dd, J=12.7, 9.5 Hz, 1H), 3.72 (q, J=7.0 Hz, 1H), 2.51 (dq, J=13.7, 6.8,6.1 Hz, 1H), 2.15 (ddd, J=14.7, 8.3, 2.3 Hz, 1H), 1.94 (d, J=12.7 Hz,1H), 1.77 (ddd, J=12.7, 4.0, 2.9 Hz, 1H), 1.61 (dt, J=14.6, 5.2 Hz, 2H),1.24 (t, J=7.0 Hz, 1H), 1.09 (d, J=7.2 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₂H₂₂F₂N₃O₅: 446.15; found: 446.2.

Step 5b

Magnesium bromide (59 mg, 0.32 mmol) was added to a solution ofintermediate 40-A (70 mg, 0.15 mmol) in 2 mL acetonitrile. The reactionmixture was stirred at 50° C. for 1 hour, acidified with 10% aqueousHCl, partitioned between the aqueous and dichloromethane, and theaqueous phase extracted to dichloromethane. The combined organic phaseswere dried over MgSO₄, filtered, concentrated, and purified by silicagel chromatography (0-10% EtOH/DCM) to afford compound 40. ¹H-NMR (400MHz, Chloroform-d) δ 12.32 (s, 1H), 10.36 (s, 1H), 8.29 (s, 1H),7.44-7.33 (m, 1H), 6.88-6.76 (m, 2H), 5.37 (dd, J=9.5, 4.1 Hz, 1H), 5.28(t, J=5.3 Hz, 1H), 4.63 (d, J=5.9 Hz, 2H), 4.23 (d, J=23.0 Hz, 2H), 3.99(dd, J=12.7, 9.5 Hz, 1H), 3.72 (q, J=7.0 Hz, 1H), 2.51 (dq, J=13.7, 6.8,6.1 Hz, 1H), 2.15 (ddd, J=14.7, 8.3, 2.3 Hz, 1H), 1.94 (d, J=12.7 Hz,1H), 1.77 (ddd, J=12.7, 4.0, 2.9 Hz, 1H), 1.61 (dt, J=14.6, 5.2 Hz, 2H),1.24 (t, J=7.0 Hz, 1H), 1.09 (d, J=7.2 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₂H₂₂F₂N₃O: 446.15; found: 446.2.

Example 41 Preparation of Compound 41(1R,4S,12aR)-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A solution of the 41-A (2020 mg, 7.463 mmol) (prepared by the samemethod as 38-A) in THF (14 mL) was stirred at 0° C. as 2.0 M LiBH₄ inTHF (7.5 mL, mmol) was added. After the resulting mixture was stirred atrt for 21 h, it was cooled at 0° C. and diluted with EA before water wasadded slowly to quench. After two phases were separated, the aqueousfraction was extracted with EA (×1) and the two organic fractions werewashed with water (×1), combined, dried (Na₂SO₄), and concentrated. Theresidue was purified by CombiFlash (120 g column) using hexanes-EA aseluents to get 41-B. LCMS-ESI⁺ (m/z): [M−C₄H₈+H]⁺ calculated forC₈H₁₄NO₃: 172.10; found: 171.95.

Step 2

A 100-mL round bottom flask was charged with reactant 41-B (1.6 g, 7.05mmol) and triethylamine (0.94 g, 9.3 mmol) in DCM (20 mL).Methanesulfonyl chloride (0.91 g, 8.0 mmol) was added to the reactionmixture. Then the reaction mixture was stirred at room temperature for 3hours. The mixture was diluted with EA (100 mL) and washed with water(2×). The aqueous fractions were extracted with EA (1×), and the organicfractions were combined, dried (Na2SO4), and concentrated. The residuewas purified by Combi Flash (120 g column, cartridge used) usinghexanes-EA as eluents to afford 41-C. LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₁₈H₁₉F₂N₂O₇: 306; found: 306.

Step 3

A 100-mL round bottom flask was charged with reactant 41-C (2.1 g, 6.9mmol) and sodium azide (2.3 g, 34.5 mmol) in DMF (10 mL). Then thereaction mixture was stirred at 100° C. for overnight. The mixture wasdiluted with EA (100 mL) and washed with water (2×). The aqueousfractions were extracted with EA (1×), and the organic fractions werecombined, dried (Na₂SO₄), and concentrated. The residue was purified byCombi Flash (120 g column, cartridge used) using hexanes-EA as eluentsto afford 41-D. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₈H₁₉F₂N₂O₇:253; found: 253.

Step 4

To a solution (purged with N₂) of reactant 41-D (1.3 g) in EA (20 mL)and EtOH (20 mL) was added Pd/C (130 mg). The mixture was stirred underH₂ for 3 hours. The mixture was filtered through celite and the filtratewas concentrated to afford compound 41-E. LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₁₈H₁₉F₂N₂O₇: 227; found: 227.

Step 5

A 100-mL round bottom flask was charged with reactant 41-E (1.05 g, 4.62mmol) and reactant 38-F (1.6 g, 4.62 mmol) in Ethanol (20 mL). Sodiumbicarbonate (0.77 g, 9.2 mmol) in water (20 mL) was added to thereaction mixture.

Then the reaction mixture was stirred at room temperature overnight. Themixture was diluted with EA (100 mL) and washed with water (2×). Theaqueous fractions were extracted with EA (1×), and the organic fractionswere combined, dried (Na2SO4), and concentrated. The crude product (2.4g) was used for next step without further purification. LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₁₈H₁₉F₂N₂O₇: 556; found: 556.

A 100-mL round bottom flask was charged with the crude product from theprevious reaction in 4 N HCl/dioxane (24.7 mL). Then the reactionmixture was stirred at room temperature for 1 hour. After concentration,the intermediate (2.1 g) and DBU (3.27 g, 21.5 mmol) in toluene (30 mL)was heated to 110° C. with stirring for 1 hour. After concentration, theresidue was purified by CombiFlash (120 g column) using hexanes-ethylacetate as eluents to afford 41-F. LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₁₈H₁₉F₂N₂O₇: 409; found: 409.

Step 6

A 100-mL round bottom flask was charged with reactant 41-F (0.5 g, 1.22mmol) in THF (5 mL) and MeOH (5 mL). 1 N KOH (3.7 mL) was added to thereaction mixture. Then the reaction mixture was stirred at roomtemperature for 1 hour. The reaction mixture was acidified by adding 1 NHCl (3.7 mL), concentrated to remove most of organic solvents, andextracted with EtOAc (2×). The organic layers were combined, dried(Na₂SO₄), and concentrated to afford compound 41-G.

Step 7

A 100-mL round bottom flask was charged with reactant 41-G (0.14 g, 0.37mmol), (2,4,6-trifluorophenyl)methanamine (0.12 g, 0.73 mmol),N,N-diisopropylethylamine (DIPEA) (0.24 g, 1.84 mmol) and HATU (0.28 g,0.74 mmol) were dissolved in DCM (5 mL). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted with EA(100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl (2×) anddried over Na₂SO₄. After concentration, the crude was purified by columnchromatography on silica gel with hexane-EtOAc to afford compound 41-H.LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₈H₁₉F₂N₂O₇: 524.5; found:524.5.

Step 8

A 50-mL round bottom flask was charged with reactant 41-H (0.13 g, 0.25mmol) in TFA (2 mL). The reaction mixture was stirred at roomtemperature for 30 minutes. After concentration, the crude was purifiedby column chromatography on silica gel with EtOAc-MeOH to affordcompound 41. ¹H-NMR (400 MHz, Chloroform-d) δ 11.61 (s, 1H), 10.70-10.01(m, 1H), 8.26 (s, 1H), 6.65 (t, J=8.1 Hz, 2H), 4.88 (s, 1H), 4.65 (dd,J=6.1, 2.4 Hz, 2H), 4.07 (d, J=10.9 Hz, 1H), 3.93-3.58 (m, 2H), 2.67 (d,J=3.1 Hz, 1H), 2.08-1.41 (m, 7H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ−109.22 (d, J=11.6 Hz, 1F), −111.04˜−112.79 (m, 2F). LCMS-ESI (m/z):[M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₅: 434; found: 434.

Example 42 Preparation of Compound 42(2R,5S,13aR)-8-hydroxy-7,9-dioxo-N-(2,4,6-trifluorobenzyl)-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

1-(2,2-dimethoxyethyl)-5-methoxy-6-(methoxycarbonyl)-4-oxo-1,4-dihydropyridine-3-carboxylicacid (3.5 g, 10 mmol) in acetonitrile (36 mL) and acetic acid (4 mL) wastreated with methanesulfonic acid (0.195 mL, 3 mmol) and placed in a 75deg C. bath. The reaction mixture was stirred for 7 h, cooled and storedat −10° C. for 3 days and reheated to 75° C. for an additional 2 h. Thismaterial was cooled and carried on crude to the next step.

Step 2

Crude reaction mixture from step 1 (20 mL, 4.9 mmol) was transferred toa flask containing (1R,3S)-3-aminocyclopentanol (0.809 g, 8 mmol). Themixture was diluted with acetonitrile (16.8 mL), treated with potassiumcarbonate (0.553 g, 4 mmol) and heated to 85° C. After 2 h, the reactionmixture was cooled to ambient temperature and stirred overnight. 0.2MHCl (50 mL) was added, and the clear yellow solution was extracted withdichloromethane (2×150 mL). The combined organic layers were dried oversodium sulfate, filtered and concentrated to 1.49 g of a light orangesolid. Recrystallization from dichlormethane:hexanes afforded thedesired intermediate 42A: LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₅H₁₇N₂O₆: 321.11; found: 321.3.

Step 3

Intermediate 42-A (0.225 g, 0.702 mmol) and(2,4,6-trifluorophenyl)methanamine (0.125 g, 0.773 mmol) were suspendedin acetonitrile (4 mL) and treated with N,N-diisopropylethylamine(DIPEA) (0.183 mmol, 1.05 mmol). To this suspension was added(dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methaniminiumhexafluorophosphate (HATU, 0.294 g, 0.774 mmol). After 1.5 hours, thecrude reaction mixture was taken on to the next step. LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₂H₂₁F₃N₃O₅: 464.14; found: 464.2.

Step 4

To the crude reaction mixture of the previous step was added MgBr₂(0.258 g, 1.40 mmol). The reaction mixture was stirred at 50° C. for 10minutes, acidified with 10% aqueous HCl, and extract twice withdichloromethane. The combined organic phases were dried over MgSO₄,filtered, concentrated, and purified by silica gel chromatography(EtOH/dichlormethane) followed by HPLC (ACN/H₂O with 0.1% TFA modifier)to afford compound 42: ¹H-NMR (400 MHz, DMSO-d₆) δ 12.43 (s, 1H), 10.34(t, J=5.7 Hz, 1H), 8.42 (s, 1H), 7.19 (t, J=8.7 Hz, 2H), 5.43 (dd,J=9.5, 4.1 Hz, 1H), 5.08 (s, 1H), 4.66 (dd, J=12.9, 4.0 Hz, 1H), 4.59(s, 1H), 4.56-4.45 (m, 2H), 4.01 (dd, J=12.7, 9.7 Hz, 1H), 1.93 (s, 4H),1.83 (d, J=12.0 Hz, 1H), 1.56 (dt, J=12.0, 3.4 Hz, 1H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₁H₁₉F₃N₃O₅: 450.13; found: 450.2.

Example 43 Preparation of Compound 43(12aR)—N—((R)-1-(2,4-difluorophenyl)ethyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL round bottom flask was charged with reactant 41-G (0.14 g, 0.37mmol), (R)-1-(2,4-difluorophenyl)ethanamine (0.12 g, 0.74 mmol),N,N-diisopropylethylamine (0.24 g, 1.84 mmol) and HATU (0.28 g, 0.74mmol) and were dissolved in DCM (5 mL). The reaction mixture was stirredat room temperature for 2 hours. The mixture was diluted with EA (100mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl (2×) anddried over Na₂SO₄. After concentration, the crude was purified by columnchromatography on silica gel with hexane-EtOAc to afford compound 43-A.LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₈H₁₉F₂N₂O₇: 520; found: 520.

Steps 2

A 50-mL round bottom flask was charged with reactant 43-A (0.14 g, 0.27mmol) in TFA (2 mL). The reaction mixture was stirred at roomtemperature for 30 minutes. After concentration, the crude was purifiedby column chromatography on silica gel with EtOAc-MeOH to affordcompound 43. ¹H-NMR (400 MHz, Chloroform-d) δ 11.65 (s, 1H), 10.57 (s,1H), 8.22 (s, 1H), 7.31 (m, 1H), 6.99-6.62 (m, 2H), 5.64-5.32 (m, 1H),4.90 (d, J=2.7 Hz, 1H), 4.04 (d, J=11.5 Hz, 1H), 3.93-3.63 (m, 2H), 2.67(s, 1H), 2.08-1.40 (m, 9H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −113.09(m, 1F), −115.01 (m, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₂₀F₂N₃O₅: 430; found: 430.

Example 44 Preparation of Compound 44(13aS)-8-hydroxy-7,9-dioxo-N-(2,3,4-trifluorobenzyl)-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

Compound 15-B (40 mg, 0.12 mmol) was taken up in 1 mL acetonitrile andtreated with 2,3,4-trifluorobenzylamine (29 mg, 0.18 mmol), HATU (53 mg,0.14 mmol), N,N-diisopropylethylamine (DIPEA) (20 mg, 0.16 mmol), andstirred at room temperature for 2 hours, after which LCMS analysisrevealed complete consumption of compound 15-B and formation ofintermediate 44-A. The reaction mixture was carried onto the next step.

Step 2

To the crude reaction solution of the previous step was added MgBr₂ (63mg, 0.34 mmol). The reaction mixture was stirred at 50° C. for one hour,acidified with 10% aqueous HCl, partitioned between the aqueous anddichloromethane, and the aqueous phase extracted to dichloromethane. Thecombined organic phases were dried over MgSO₄, filtered, concentrated,and purified by HPLC (ACN/H₂O with 0.1% TFA modifier) to compound 44.¹H-NMR (400 MHz, DMSO-d₆) δ 12.45 (s, 1H), 10.38 (t, J=6.0 Hz, 1H), 8.43(s, 1H), 7.27 (q, J=9.2 Hz, 1H), 7.16 (q, J=8.5 Hz, 1H), 5.42 (dd,J=9.5, 4.0 Hz, 1H), 5.08 (s, 1H), 4.76-4.47 (m, 4H), 4.01 (dd, J=12.8,9.7 Hz, 1H), 1.92 (s, 4H), 1.82 (d, J=12.1 Hz, 1H), 1.55 (dt, J=12.2,2.9 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₉F₃N₃O₅:450.13; found: 450.2.

Example 45 Preparation of Compound 45(13aS)-8-hydroxy-7,9-dioxo-N-(2,4,6-trifluorobenzyl)-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

Compound 15-B (38 mg, 0.12 mmol) was taken up in 1 mL acetonitrile andtreated with 2,4,6-trifluorobenzylamine (34 mg, 0.21 mmol), HATU (50 mg,0.13 mmol), N,N-diisopropylethylamine (DIPEA) (23 mg, 0.18 mmol), andstirred at room temperature for 2 hours, after which LCMS analysisrevealed complete consumption of compound 15-B and formation ofintermediate 45-A. The reaction mixture was carried onto the next step.

Step 2

To the crude reaction solution of the previous step was added MgBr₂ (55mg, 0.30 mmol). The reaction mixture was stirred at 50° C. for one hour,acidified with 10% aqueous HCl, partitioned between the aqueous anddichloromethane, and the aqueous phase extracted with dichloromethane.The combined organic phases were dried over MgSO₄, filtered,concentrated, and purified by HPLC (ACN/H₂O with 0.1% TFA modifier) toafford compound 45. ¹H-NMR (400 MHz, DMSO-d₆) δ 12.37 (s, 1H),10.37-10.25 (m, 1H), 8.37 (s, 1H), 7.14 (t, J=8.7 Hz, 2H), 5.37 (dd,J=9.5, 4.0 Hz, 1H), 5.02 (s, 1H), 4.66-4.40 (m, 4H), 3.95 (dd, J=12.8,9.6 Hz, 1H), 1.87 (s, 4H), 1.77 (d, J=11.9 Hz, 1H), 1.50 (dt, J=11.8,3.2 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₉F₃N₃O₅:450.13; found: 450.2.

Example 46 Preparation of Compound 46(13aS)—N-(2,6-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

Compound 15-B (38 mg, 0.12 mmol) was taken up in 1 mL acetonitrile andtreated with 2,6-difluorobenzylamine (19 mg, 0.14 mmol), HATU (56 mg,0.15 mmol), N,N-diisopropylethylamine (DIPEA) (20 mg, 0.15 mmol), andstirred at room temperature for 90 minutes, after which LCMS analysisrevealed complete consumption of compound A and formation ofintermediate 46-A. The reaction mixture was carried onto the next step.

Step 2

To the crude reaction solution of the previous step was added MgBr₂ (50mg, 0.27 mmol). The reaction mixture was stirred at 50° C. for one hour,acidified with 10% aqueous HCl, partitioned between the aqueous anddichloromethane, and the aqueous phase extracted with dichloromethane.The combined organic phases were dried over MgSO₄, filtered,concentrated, and purified by HPLC (ACN/H₂O with 0.1% TFA modifier) toafford compound 46. ¹H-NMR (400 MHz, DMSO-d₆) δ 12.37 (s, 1H),10.33-10.26 (m, 1H), 8.37 (s, 1H), 7.39-7.29 (m, 1H), 7.05 (t, J=7.9 Hz,2H), 5.37 (dd, J=9.5, 4.1 Hz, 1H), 5.02 (s, 1H), 4.66-4.45 (m, 4H), 3.95(dd, J=12.7, 9.6 Hz, 1H), 1.87 (s, 4H), 1.77 (d, J=12.0 Hz, 1H), 1.50(dt, J=12.2, 3.5 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₂₀F₂N₃O₅: 432.14; found: 432.2.

Example 47 Preparation of Compound 47(1R,4S,12aR)—N-(2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

The crude acid 41-G (0.45 g, 1.18 mmol), 2,4-difluobenzylamine (0.35 g,2.44 mmol), N,N-diisopropylethylamine (DIPEA) (0.79 g, 6.11 mmol) andHATU (0.93 g, 2.44 mmol) were dissolved in DCM (10 mL). The reactionmixture was stirred at room temperature for 2 hours. The mixture wasdiluted with EA (100 mL) and washed with saturated NaHCO₃ (2×),saturated NH₄Cl (2×) and dried over Na₂SO₄. After concentration, thecrude was purified by column chromatography on silica gel withhexane-EtOAc to afford compound 47-A. LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₁₈H₁₉F₂N₂O₇: 506; found: 506.

Step 2

A 50-mL round bottom flask was charged with reactant 47-A (0.5 g, 0.99mmol) in TFA (6 mL). The reaction mixture was stirred at roomtemperature for 30 minutes. After concentration, the crude was purifiedby column chromatography on silica gel with EtOAc-MeOH to affordcompound 47. ¹H NMR (400 MHz, Chloroform-d) δ 11.70 (s, 1H), 10.44 (s,1H), 8.29 (s, 1H), 7.60-7.29 (m, 1H), 6.95-6.58 (m, 2H), 4.10 (s, 1H),4.02-3.54 (m, 3H), 2.68 (d, J=3.1 Hz, 1H), 2.00-1.40 (m, 8H). ¹⁹F NMR(376 MHz, Chloroform-d) δ −112.31 (d, J=8.0 Hz, 1F), −114.77 (d, J=8.4Hz, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₅: 416;found: 416.

Example 48 Preparation of Compound 48(1S,4R,12aS)—N-(2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

48-B was prepared analogously to 55-H in Example 55, substituting 48-Afor 55-A. Compound 48 was prepared as described for compound 38 inExample 38, substituting 48-B for 38-B to afford compound 48. ¹H-NMR(400 MHz, Chloroform-d) δ 11.79 (s, 1H), 10.44 (m, 1H), 8.33 (s, 1H),7.42-7.31 (m, 1H), 6.86-6.74 (m, 2H), 4.74 (s, 1H), 4.63 (d, J=5.8 Hz,2H), 4.19 (m, 1H), 4.07-4.03 (m, 2H), 2.83 (s, 1H), 1.92-1.68 (m, 6H).¹⁹F NMR (376 MHz, Chloroform-d) δ −112.3 (m, 1F), −114.8 (m, 1F).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₄: 416.14; found:416.07.

Example 49 Preparation of Compound 49(2S,5R,13aS)-8-hydroxy-7,9-dioxo-N-((3-(trifluoromethyl)pyridin-2-yl)methyl)-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

Compound 15-B (44 mg, 0.14 mmol) was taken up in 1 mL acetonitrile andtreated with (3-(trifluoromethyl)pyridin-2-yl)methanamine (38 mg, 0.18mmol, HCl salt), HATU (69 mg, 0.18 mmol), N,N-diisopropylethylamine(DIPEA) (0.07 mL, 0.40 mmol), and stirred at room temperature for 1hour, after which LCMS analysis revealed complete consumption ofcompound 15-B and formation of intermediate 49-A. The reaction mixturewas carried onto the next step.

Step 2

To the crude reaction solution of the previous step was added MgBr₂ (51mg, 0.28 mmol). The reaction mixture was stirred at 50° C. for 90minutes, acidified with 10% aqueous HCl, partitioned between the aqueousand dichloromethane, and the aqueous phase extracted withdichloromethane. The combined organic phases were dried over MgSO₄,filtered, concentrated, and triturated by methanol followed by diethylether to afford compound 49. ¹H-NMR (400 MHz, DMSO-d₆) δ 12.42 (s, 1H),10.80-10.70 (m, 1H), 8.83 (d, J=5.0 Hz, 1H), 8.44 (s, 1H), 8.19 (d,J=8.6 Hz, 1H), 7.56 (dd, J=7.7, 5.2 Hz, 1H), 5.43 (dd, J=9.5, 4.0 Hz,1H), 5.08 (s, 1H), 4.86-4.80 (m, 2H), 4.67 (dd, J=12.9, 4.0 Hz, 1H),4.59 (s, 1H), 4.02 (dd, J=12.6, 9.8 Hz, 1H), 1.93 (s, 4H), 1.82 (d,J=12.1 Hz, 1H), 1.60-1.52 (m, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₁H₂₀F₃N₄O₅: 465.14; found: 465.2.

Examples 50 and 51 Preparation of Compounds 50 and 51N-(2,4-difluorobenzyl)-9-hydroxy-8,10-dioxo-2,3,5,6,8,10,14,14a-octahydro-2,6-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,6,3]dioxazocine-11-carboxamide50 and 51

Step 1

Methyl5-(2,4-difluorobenzylcarbamoyl)-1-(2,2-dihydroxyethyl)-3-methoxy-4-oxo-1,4-dihydropyridine-2-carboxylate(1-C, 392 mg, 0.95 mmol) (Example 87), racemiccis-5-aminotetrahydro-2H-pyran-3-ol (WO 2012/145569 Bennett, B. L. etal, filed Apr. 20, 2012) (112 mg, 0.95 mmol), and potassium carbonate(134 mg, 0.97 mmol) were taken up in 3.8 mL acetonitrile/0.2 mL aceticacid and stirred at 90° C. for 90 minutes, after which the reactionmixture was partitioned between DCM and brine, the aqueous phaseextracted with DCM, combined organic phases dried over MgSO₄, filtered,concentrated, and purified by SGC (0-10% EtOH/DCM) to affordintermediate 50-A.

Step 2

Intermediate 50-A (40 mg) was separated by chiral SFC on a Chiralpak ICcolumn using 10% DMF in supercritical carbon dioxide as eluent to affordIntermediates 50-B (first eluting peak) and 51-A (second eluting peak)in enantioenriched form. For intermediate 50-B: (absolutestereochemistry unknown), Chiral HPLC retention time=11.48 minutes(Chiralpak IC, 150×4.6 mm, 1 mL/min MeOH). For intermediate 51-A:(absolute stereochemistry unknown), Chiral HPLC retention time=14.35minutes (Chiralpak IC, 150×4.6 mm, 1 mL/min MeOH).

Step 3a

Magnesium bromide (12 mg, 0.06 mmol) was added to a solution ofintermediate 50-B (10.5 mg, 0.02 mmol, absolute stereochemistry unknown)in 1 mL acetonitrile. The reaction mixture was stirred at 50° C. for 1hour, acidified with 10% aqueous HCl, partitioned between the aqueousand dichloromethane, and the aqueous phase extracted withdichloromethane. The combined organic phases were dried over MgSO₄,filtered, concentrated, and purified by HPLC (ACN/H₂O with 0.1% TFAmodifier) to afford compound 50. ¹H-NMR (400 MHz, Chloroform-d) δ 10.47(t, J=5.8 Hz, 1H), 8.42 (s, 1H), 7.35 (q, J=8.6, 8.2 Hz, 1H), 6.81 (q,0.1=8.7, 8.0 Hz, 2H), 6.41 (dd, J=10.0, 3.6 Hz, 1H), 4.79 (s, 1H), 4.65(s, 2H), 4.36-4.26 (m, 2H), 4.20-4.08 (m, 2H), 3.98 (dd, J=12.4, 10.2Hz, 1H), 3.88 (t, J=11.8 Hz, 2H), 2.27 (dt, J=13.3, 3.1 Hz, 1H),2.15-2.06 (m, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₆:448.40; found: 448.2.

Step 3b

Magnesium bromide (13 mg, 0.07 mmol) was added to a solution ofintermediate 51-A (13.2 mg, 0.03 mmol, absolute stereochemistry unknown)in 1 mL acetonitrile. The reaction mixture was stirred at 50° C. for 1hour, acidified with 10%° aqueous HCl, partitioned between the aqueousand dichloromethane, and the aqueous phase extracted withdichloromethane. The combined organic phases were dried over MgSO₄,filtered, concentrated, and purified by HPLC (ACN/H₂O with 0.1% TFAmodifier) to afford compound 51. ¹H-NMR (400 MHz, Chloroform-d) δ 10.47(t, J=5.8 Hz, 1H), 8.42 (s, 1H), 7.35 (q, J=8.6, 8.2 Hz, 1H), 6.81 (q,J=8.7, 8.0 Hz, 2H), 6.41 (dd, J=10.0, 3.6 Hz, 1H), 4.79 (s, 1H), 4.65(s, 2H), 4.36-4.26 (m, 2H), 4.20-4.08 (m, 2H), 3.98 (dd, J=12.4, 10.2Hz, 1H), 3.88 (t, J=11.8 Hz, 2H), 2.27 (dt, J=13.3, 3.1 Hz, 1H),2.15-2.06 (m, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₆:448.40; found: 448.2.

Example 52 Preparation of Compound 52(2S,5R,13aS)—N-(2-cyclopropoxy-4-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

A solution cyclopropanol (1.9 g, 29 mmol) in 20 mL dioxane was addeddropwise to a 0° C. solution of Sodium hydride (60% dispersion inmineral oil, 1.04 g, 26 mmol) in 80 mL dioxane. The reaction mixture wasallowed to warm to room temperature, 2,4-difluorobenzonitrile (3.48 g,25 mmol) was added portionwise, and reaction temperature raised to 95°C. The reaction solution was cooled to room temperature after stirringfor 18 hours, diluted with ethyl acetate, washed twice with water andtwice with brine, dried over MgSO₄, filtered, and concentrated ontosilica gel. Purification by silica gel chromatography (0-10%EtOAc/hexanes) afforded 2-cyclopropoxy-4-fluorobenzonitrile. ¹H-NMR (400MHz, Chloroform-d) δ 7.52 (dd, J=8.6, 6.2 Hz, 1H), 7.05 (dd, J=10.5, 2.3Hz, 1H), 6.73 (td, J=8.2, 2.3 Hz, 1H), 3.87-3.76 (m, 1H), 0.87 (m, 4H).

Step 2

To a 0° C. suspension of lithium aluminum hydride in THF (1M, 15 mL,mmol) was added 2-cyclopropoxy-4-fluorobenzonitrile in 14 mL diethylether dropwise. The reaction solution was stirred for 3 hours, graduallywarming to room temperature, at which point it was recooled to 0° C., anadditional 8 mL lithium aluminum hydride in THF (1M, 8 mmol) added, andstirred for an additional 90 minutes. The reaction was quenched bysequential addition of 0.9 mL water, 0.9 mL 15% NaOH_((aq)), and 2.7 mLwater. The reaction was filtered through celite with diethyl etherrinses, dried over MgSO₄, and concentrated to afford2-cyclopropoxy-4-fluorobenzylamine of sufficient purity to carry on ascrude. ¹H-NMR (400 MHz, Chloroform-d) δ 7.17-7.08 (m, 1H), 6.96 (dd,J=10.9, 2.4 Hz, 1H), 6.61 (td. J=8.3, 2.5 Hz, 1H), 3.78-3.66 (m, 3H),0.89-0.72 (m, 4H).

Step 3

Compound 15-B (46 mg, 0.14 mmol) was taken up in 1 mL acetonitrile andtreated with 2-cyclopropoxy-4-fluorobenzylamine (32 mg, 0.18 mmol), HATU(62 mg, 0.16 mmol), N,N-diisopropylethylamine (DIPEA) (0.04 mL, 0.22mmol), and stirred at room temperature for 2 hours, after which LCMSanalysis revealed complete consumption of compound 15-B and formation ofintermediate 52-A. The reaction mixture was carried onto the next step.

Step 4

To the crude reaction solution of the previous step was added MgBr₂ (56mg, 0.30 mmol). The reaction mixture was stirred at 50° C. for 90minutes, acidified with 10% aqueous HCl, partitioned between the aqueousand dichloromethane, and the aqueous phase extracted withdichloromethane. The combined organic phases were dried over MgSO₄,filtered, concentrated, and purified by HPLC (ACN/H₂O with 0.1% TFAmodifier) to afford compound 52. ¹H-NMR (400 MHz, DMSO-d₆) δ 12.44 (s,1H), 10.21 (t, J=5.8 Hz, 1H), 8.41 (s, 1H), 7.22-7.15 (m, 1H), 7.12 (dd,J=11.2, 2.5 Hz, 1H), 6.72 (td, J=8.5, 2.5 Hz, 1H), 5.42 (dd, J=9.6, 4.1Hz, 1H), 5.07 (s, 1H), 4.66 (dd, J=12.8, 4.1 Hz, 1H), 4.58 (s, 1H), 4.34(dd, J=5.6, 2.4 Hz, 2H), 4.04-3.91 (m, 2H), 1.92 (s, 4H), 1.82 (d,J=11.9 Hz, 1H), 1.55 (dt, J=12.4, 3.5 Hz, 1H), 0.80 (q, J=6.3, 5.7 Hz,2H), 0.72 (q, J=6.0, 4.9 Hz, 2H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₄H₂₅FN₃O₆: 470.17; found: 470.1.

Example 53 Preparation of Compound 53(2R,5S,13aR)—N-(2-cyclopropoxy-4-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

Compound 42-A (46 mg, 0.14 mmol) was taken up in 1 mL acetonitrile andtreated with 2-cyclopropoxy-4-fluorobenzylamine (33 mg, 0.18 mmol), HATU(61 mg, 0.16 mmol), N,N-diisopropylethylamine (DIPEA) (0.04 mL, 0.24mmol), and stirred at room temperature for 2 hours, after which LCMSanalysis revealed complete consumption of compound 42-A and formation ofintermediate 53-A. The reaction mixture was carried onto the next step.

Step 2

To the crude reaction solution of the previous step was added MgBr₂ (55mg, 0.30 mmol). The reaction mixture was stirred at 50° C. for 90minutes, acidified with 10% aqueous HCl, partitioned between the aqueousand dichloromethane, and the aqueous phase extracted withdichloromethane. The combined organic phases were dried over MgSO₄,filtered, concentrated, and purified by HPLC (ACN/H₂O with 0.1% TFAmodifier) to afford compound 53. ¹H-NMR (400 MHz, DMSO-d₆) δ 12.44 (s,1H), 10.21 (t, J=5.8 Hz, 1H), 8.41 (s, 1H), 7.22-7.15 (m, 1H), 7.12 (dd,J=11.2, 2.5 Hz, 1H), 6.72 (td, J=8.5, 2.5 Hz, 1H), 5.42 (dd, J=9.6, 4.1Hz, 1H), 5.07 (s, 1H), 4.66 (dd, J=12.8, 4.1 Hz, 1H), 4.58 (s, 1H), 4.34(dd, J=5.6, 2.4 Hz, 2H), 4.04-3.91 (m, 2H), 1.92 (s, 4H), 1.82 (d,J=11.9 Hz, 1H), 1.55 (dt, J=12.4, 3.5 Hz, 1H), 0.80 (q, J=6.3, 5.7 Hz,2H), 0.72 (q, J=6.0, 4.9 Hz, 2H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₄H₂₅FN₃O₆: 470.17; found: 470.1.

Example 54 Preparation of Compound 54(2R,5S)—N—((S)-1-(2,4-difluorophenyl)-2,2,2-trifluoroethyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

A 50-mL round bottom flask was charged with reactant 54-A (0.02 g, 0.06mmol), (S)-1-(2,4-difluorophenyl)-2,2,2-trifluoroethanamine (0.019 g,0.09 mmol), N,N-diisopropylethylamine (DIPEA) (0.048 g, 0.38 mmol) andHATU (0.036 g, 0.09 mmol) in DCM (2 ml). The reaction mixture wasstirred at room temperature for 1 hour. The reaction mixture wasconcentrated down, re-dissolved in EtOAc (50 mL), washed with saturatedNaHCO₃ (2×), saturated NH₄Cl and dried over Na₂SO₄. After concentration,the crude was purified by column chromatography on silica gel withhexane-EtOAc to obtain 54-B. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₈H₁₉F₂N₂O₇: 514; found: 514.

Step 2

A 50-mL round bottom flask was charged with reactant 54-B (0.03 g, 0.058mmol) and magnesium bromide (0.03 g, 0.15 mmol) in acetonitrile (2 mL).The reaction mixture was heated to 50° C. After 10 minutes, the reactionmixture was cooled to 0° C. and 1 N hydrochloric acid (0.5 mL) was addedin. Then the reaction mixture was diluted with MeOH (2 mL). Afterfiltration, the crude was purified by Pre-HPLC purification (30-70%acetonitrile:water, 0.1% TFA) afforded compound 54 as TFA salt. ¹H-NMR(400 MHz, Chloroform-d) δ 11.28 (d, J=9.4 Hz, 1H), 8.39 (s, 1H), 7.54(q, J=7.8 Hz, 1H), 7.12-6.76 (m, 2H), 6.40-5.98 (m, 1H), 5.57-5.18 (m,2H), 4.68 (s, 1H), 4.29 (dd, J=13.1, 4.0 Hz, 1H), 4.05 (dd, J=12.9, 9.3Hz, 1H), 2.39-1.94 (m, 4H), 1.86 (t, J=10.5 Hz, 1H), 1.60 (dt, J=12.6,3.4 Hz, 1H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −75.30 (t, J=6.8 Hz, 3F),−108.33 (dd, J=8.6, 6.3 Hz, 1F), −111.56-−113.23 (m, 1F). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₅: 500; found: 500.

Example 55 Preparation of Compound 55(1R,4S,12aS)-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A mixture of compound 55-A (40.60 g, 150 mmol) and Pd(OH)₂/C (12 g) inEtOH (400 mL) under an atmosphere of H₂ was stirred at room temperatureovernight. The reaction mixture was filtered and treated with HCl/EtOH(400 ml). The mixture was stirred at room temperature for 2 h. Thereaction mixture was concentrated to give compound 55-B, which was usedin next step without purification. LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₉H₁₆NO: 170.1; found: 170.2.

Step 2

To a solution of compound 55-B (92.25 g, 0.45 mol) and K₂CO₃ (186.30 g,1.35 mol) in CH₃CN (1 L) was added benzyl bromide (76.50 g, 0.45 mol) at0° C. The mixture was stirred at room temperature overnight. Thereaction mixture was filtered, concentrated and the residue was purifiedby chromatography on silica gel to give compound 55-C.

Step 3

To a mixture of diisopropylamine (50 g, 0.50 mol) in THF (400 mL) wasadded n-BuLi (200 mL, 0.50 mol) at −78° C. at N₂ atmosphere. After 0.5h, the reaction mixture was warmed to 20° C. and stirred for 0.5 h. Themixture was cooled to −78° C. and added a solution of compound 55-C(64.75 g, 0.25 mol) in THF (600 mL) under N₂ atmosphere. The mixture wasstirred for 4 h and quenched with saturated NH₄Cl solution. The mixturewas extracted with EtOAc and the organic layer was washed with brine,dried over Na₂SO₄, filtered and concentrated. The residue was purifiedby chromatography on silica gel to give compound 55-D.

Step 4

A mixture of compound 55-D (129.50 g 0.50 mol) in 4N HCl (1.30 L) wasrefluxed for 4 h. the mixture was concentrated. The residue was purifiedby HPLC to give compound 55-E.

Step 5

To a mixture of compound 55-E (47 g, 176 mmol) and Pd(OH)₂/C (9 g) inEtOH (400 mL) under an atmosphere of H₂ was stirred at room temperatureovernight. The reaction mixture was concentrated to give compound 55-F,which was used in next step without purification. ¹H-NMR (400 MHz,CDCl₃) δ 4.22 (s, 1H), 4.06 (s, 1H), 2.98-2.95 (d, J=11.2 Hz, 1H),1.96-1.93 (d, J=11.2 Hz, 1H), 1.86-1.82 (m, 2H), 1.76-1.74 (d, J=9.2 Hz,2H), 1.49 (s, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₇H₁₂NO₂:142.1; found: 142.1.

Step 6

To a mixture of compound 55-F (29.20 g, 165 mmol) and 2N NaOH solution(330 mL, 0.66 mol) in dioxane (120 mL) was added Boc₂O (39.60 g, 181mmol) at 0° C. The reaction mixture was stirred at room temperatureovernight. The mixture was adjusted with 3N HCl to pH=5-6 and extractedwith DCM. The organic layer was dried over Na₂SO₄, filtered andconcentrated to give 55-G. ¹H-NMR (400 MHz, CDCl₃) δ 4.40 (s, 1H), 4.26(s, 1H), 2.89 (s, 1H), 1.76-1.74 (s, 1H), 1.69-1.59 (m, 4H), 1.50 (s,1H), 1.47 (s, 9H). LCMS-ESI⁺ (m/z): [M+Na]⁺ calculated for C₁₂H₁₉NNaO₄:264.1; found: 264.1.

Step 7

To a mixture of compound 55-G (500 mg, 2.07 mmol) in THF (10 mL) chilledto 0° C. was added BH₃-DMS THF complex (2N in THF, 8.23 mmol, 4.1 mL)slowly. Gas evolution occurred. Internal temperature was monitored toensure no major exotherm. Reaction was allowed to warm to r.t.overnight. Some starting material remained by LC/MS, additional 2 mLBH₃-DMS THF complex was added and the mixture was stirred for additional3 hr then cooled reaction to 0° C. and slowly quenched with methanol(gas evolution occurs). Internal temperature monitored to ensureexotherm below 25° C. The mixture was concentrated then purified bysilica gel chromotography (20-40% EtOAc/Hexanes) to afford 55-H.

Step 8

Compound 55 was prepared as described for Example 41, substituting 55-Hfor 41-B to afford compound 55. ¹H-NMR (400 MHz, DMSO-d6) δ 11.81 (s,1H), 10.40 (t, J=5.8 Hz, 1H), 8.39 (s, 1H), 7.19 (t, J=8.6 Hz, 2H),4.59-4.48 (m, 4H), 4.16 (t, J=12.2 Hz, 1H), 4.03 (d, J=12.2 Hz, 1H),2.69 (s, 1H), 1.75 (d, J=10.1 Hz, 1H), 1.69-1.55 (m, 5H). ¹⁹F NMR (376MHz, DMSO-d6) δ −109.3 (m, 1F), −112.5 (m, 1F). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₁H₁₉F₃N₃O₄: 434.13; found: 434.32.

Example 56 Preparation of Compound 56(1R,2S,4R,12aR)-2-fluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A solution of 56-A (5 g, 19.43 mmol) in tetrahydrofuran (65 ml) wascooled in an ice bath as 0.5 M 9-borabicyclo[3.3.1]nonane (48.58 ml) wasadded dropwise. The reaction mixture was warmed up to room temperature.After 18 hours, the reaction was cooled to 0° C. and a mixture of 2Msodium hydroxide (34 ml) and hydrogen peroxide (9.34 ml, 97.15 mmol) wasadded dropwise. After 2 hours at 0° C., the reaction was warmed up toroom temperature and stirred for 1 hour. The mixture was diluted withEtOAc and washed with water. The aqueous fractions were extracted withEtOAc, and the organic fractions combined were dried (Na₂SO₄) andconcentrated. The residue was purified by silica column chromatography(50-70% EtOAc/hexanes) to afford 56-B (3.05 g, 57%). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₁₆H₂₁NO₃: 275.34; found: 276.122.

Step 2

To a solution of 56-B (1.45 g, 5.27 mmol) in N,N-dimethylformamide (12ml) was added tert-butylchlorodiphenylsilane (1.51 ml, 5.79 mmol) andimidazole (1.08 g, 15.8 mmol). After 18 hours, the mixture was dilutedwith water, extracted into EtOAc (2×), the organic fractions werecombined, dried (Na₂SO₄), and concentrated. The residue was purified bysilica column chromatography (10-20% EtOAc/hexanes) to afford 56-C (2.6g, 96.1%). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₃₂H₃₉NO₃Si: 513.74;found: 514.625.

Step 3

To a solution of 56-C (3.27 g, 6.36 mmol) in EtOH (26 mL) and aceticacid (3 mL) was added 10% PdOH/C (0.52 g, 3.7 mmol) and the suspensionwas shaken in a Parr apparatus at 50 atm for 20 hours. After filteringthrough Celite, the cake was washed with EtOH, the filtrate wasconcentrated under vacuum. The residue was dissolved in ethanol (26 ml)and acetic acid (3 ml, 52.4 mmol), treated with 10% PdOH/C (0.52 g, 3.7mmol) and shaken in a Parr apparatus at 50 atm for 20 hours. Filteredthrough Celite, the cake was washed with EtOH, the filtrate wasconcentrated under vacuum to dryness to afford the crude deprotectedproduct (2.07 g, 79.4%). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₄H₃₁NO₃Si: 409.59; found: 410.485.

To the crude residue (2 g, 4.88 mmol) and di-tert-butyl dicarbonate 97%(2.14 g, 9.79 mmol) in THF (20 ml) was added N,N-diisopropylethylamine(DIPEA) (2.14 ml, 12.27 mmol). After 20 h, the reaction mixture wasdiluted with water, extracted into EtOAC (2×) and the two organicfractions were washed with water, combined, dried (Na₂SO₄), andconcentrated. The residue was purified by silica column chromatography(10-20%. EtOAc/Hexanes) to afford 56-D (2.13 g, 86.14%). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₃₀H₄₁NO₅Si: 523.74; found: 523.922.

Step 4

A solution of 56-D (2.07 g, 4.06 mmol) in THF (20 ml) was stirred in anice bath as 2.0 M LiBH₄ in THF (4.07 ml) was added and the resultingmixture was stirred at room temperature for 18 h. After, the reactionmixture was diluted with ethyl acetate and treated slowly with water.The two phases were separated, and the aqueous fraction was extractedagain with ethyl acetate. The two organic fractions were washed withwater, combined, dried (Na₂SO₄), and concentrated. The residue waspurified by silica column chromatography (20-40% EOAc/hexanes) to afford56-E (1.59 g, 81.3%). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₅H₃₉NO₄Si: 481.7; found: 482.337.

Step 5

A mixture of 56-E (1.58 g, 3.28 mmol), phthalimide (0.79 g, 5.38 mmol)and triphenylphosphine (1.93 g, 7.37 mmol) in THF (90 ml) was cooled inan ice bath. Diisopropyl azodicarboxylate, 95% (1.46 ml, 7.42 mmol) wasadded. The mixture was then warmed up to room temperature and stirredfor 20 h. After, the reaction mixture was concentrated and the residuedissolved in ether, cooled in an ice bath and stirred for 1.5 h. Thesolids were filtered off and the filtrate was concentrated. The residuewas purified by silica column chromatography (10-30/o EtOAc/hexanes) toafford the protected amino compound (1.86 g, 92.8%).

A solution of the protected amino compound 56-F (1.85 g, 3.03 mmol) andhydrazine hydrate (0.6 ml, 12.39 mmol) in ethanol (19 ml) was stirred at70° C. for 2 h. The reaction mixture was cooled in an ice bath, ether(10 ml) was added and the mixture was stirred for 30 min. The solidformed was filtered off and the filtrate was concentrated under vacuumto dryness.

Step 6

A mixture of crude amino compound 56-F (991 mg, 2.06 mmol), compound38-F (Example 38) (714 mg, 2.06 mmol) and NaHCO₃ (347 mg, 4.12 mmol) inwater (15 mL) and EtOH (15 mL) was stirred for 20 h. The reactionmixture was concentrated under vacuum and the residue was partitionedbetween water and EtOAc. The aqueous layer was re-extracted with EtOAcand the combined organic layers were dried (Na₂SO₄) and concentrated.The residue (1.5 g) was dissolved in CH₂Cl₂ (5 mL) and 4N HCl in dioxane(18.6 mL) was added. After 1.5 hours the mixture was concentrated todryness, co-evaporated with toluene and dried in vacuo.

The crude residue (1.38 g) and DBU (1.4 ml, 9.38 mmol) in toluene (25ml) was stirred at 110° C. After 35 minutes the mixture was concentratedand the residue was purified by silica column chromatography (5-15%MeOH/EtOAc) to afford 56-G (450 mg, 72.3%). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₃₉H₄₂N₂O₆Si: 662.85; found: 663.766.

Step 7

The mixture of 56-G (890 mg, 1.34 mmol) in MeOH (14 ml) and THF (14 ml)was stirred at room temperature as 1M KOH (7.09 ml) was added. After 30min the reaction mixture was neutralized with 1N HCl, extracted intoEtOAc (2×) and the combined organic extracts were dried (Na₂SO₄) andconcentrated.

A suspension of the crude residue (850 mg), 2,4,6-trifluorobenzylamine(248 mg, 1.54 mmol) and HATU (662 mg, 1.74 mmol) in dichloromethane (5ml) was stirred at room temperature as N,N-diisopropylethylamine (DIPEA)(1.63 ml, 9.37 mmol) was added. After 1 h, additional2,4,6-difluorobenzylamine (32 mg, 0.2 mmol), HATU (153 mg, 0.4 mmol) andN,N-diisopropylethylamine (DIPEA) (0.12 ml, 0.67 mmol) were added. After30 minutes the mixture was diluted with water, extracted into EtOAc (3×)the combined organic phases were dried (Na₂SO₄), concentrated and theresidue was purified by silica column chromatography (50-75%EtOAc/hexanes) to afford 56-H (919 mg, 88.23%). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₄₄H₄₂F₃N₃O₅Si: 777.9; found: 778.409.

Step 8

A solution of 56-H (915 mg, 1.18 mmol) in THF (5 ml) was stirred in anice bath as 1.0 M tetrabutylammonium fluoride in THF (1.18 ml) was addeddropwise. The resulting mixture was stirred at room temperature for 30min. The reaction mixture was concentrated under vacuum and the residuewas diluted with EtOAc, washed with water, dried (Na₂SO₄), concentratedand the residue was purified by silica column chromatography (50-75%EtOAc/hexanes then 5% MeOH/EtOAc). The resulting material (248 mg, 0.46mmol) was dissolved in dichloromethane (2 ml) cooled to −78° C. asdiethylaminosulfur trifluoride (0.07 mL, 0.55 mmol) was added dropwiseand the reaction was warmed to room temperature and stirred for 1 h. Thereaction was cooled in an ice bath and quenched with saturated NaHCO₃,two phases were separated, and the separated aqueous fraction wasextracted with CH₂Cl₂. The two organic fractions were combined dried(Na₂SO₄), and concentrated. The residue was purified by silica columnchromatography (1% MeOH/EtOAc) to afford 56-J (75 mg) (LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₈H₂₃F₄N₃O₄: 541.49; found: 542.320) and 56-I (30mg) (LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₈H₂₂F₃N₃O₄: 521.49; found:522.05).

Step 9

Compound 56-J (75 mg, 139 mmol) was dissolved in TFA (1 mL), stirred atroom temperature for 10 minutes, and the solution was concentrated. Theresidue was purified by reverse phase HPLC (Gemini, 15 to 43%ACN/H₂O+0.1% TFA) to afford compound 56. ¹H-NMR (400 MHz, DMSO-d6) δ10.67 (s, 1H), 7.80 (s, 1H), 7.17 (t, J=8.6 Hz, 2H), 5.45-5.18 (m, 1H),4.70-4.39 (m, 3H), 4.23 (d, J=11.5 Hz, 1H), 4.11-3.85 (m, 2H), 2.85 (dd,J=4.2, 2.0 Hz, 1H), 2.34-2.13 (m, 1H), 1.81 (s, 1H), 1.55-1.33 (m, 2H).¹⁹F-NMR (376 MHz, DMSO-d₆) δ 5-74.20 (m), −106.95˜−116.45 (m),−190.65˜−194.54 (m).

Example 57 Preparation of Compound 57(1R,4R,12aR)-2,2-difluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A solution of 57-A (1.45 g, 5.34 mmol) in dichloromethane (30 ml) wascooled in an ice bath as Dess Martin periodinane (4.53 g, 10.69 mmol)was added in portions and the reaction was stirred at room temperaturefor 18 h. The reaction was quenched by addition of water, theprecipitate was filtered off and a saturated solution of Na₂S₂O₃ wasadded. The mixture was stirred until the biphasic solution turned thensaturated NaHCO₃ was added and the aqueous layer extracted with CH₂Cl₂.The combined organic fractions were dried (Na₂SO₄) and concentrated. Theresidue was purified by silica column chromatography (30-50%EtOAc/Hexanes) to afford 57-B (1.13 g, 78.2%). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₁₃H₁₉NO₅: 269.29; found: 269.722.

Step 2

A solution of 57-B (0.5 g, 1.86 mmol) in dichloromethane (10 ml) wascooled to −78° C. as diethylaminosulfur trifluoride (0.52 mL, 3.91 mmol)was added dropwise and the reaction was warmed to room temperature andstirred for 18 h. The reaction was cooled in an ice bath and quenchedwith saturated NaHCO₃, two phases were separated, and the separatedaqueous fraction was extracted with CH₂Cl₂. The two organic fractionswere combined, dried (Na₂SO₄) and concentrated. The residue was purifiedby silica column chromatography (20-50% EtOAc/hexanes) to afford 57-C(518 mg, 95.39%). ¹H-NMR (400 MHz, Chloroform-d) δ 4.43 (s, 1H),4.36-4.27 (m, 1H), 4.22 (s, 1H), 3.75 (s, 3H), 2.95 (t, J=8.1 Hz, 1H),2.30-1.98 (m, 2H), 1.85-1.71 (m, 1H), 1.44 (m, 9H).

Step 3

A solution of 57-C (935 mg, 3.21 mmol) in THF (10 ml) was stirred in anice bath as 2.0 M LiBH4 in THF (3.22 ml) was added and the resultingmixture was stirred at room temperature for 18 h. After, the reactionmixture was diluted with ethyl acetate and water was added slowly. Thetwo phases were separated, and the separated aqueous fraction wasextracted with ethyl acetate. The two organic fractions were washed withwater, combined, dried (Na₂SO₄), and concentrated. The residue waspurified by silica column chromatography (20-40° % EtOAc/hexanes) toafford 57-D (724 mg, 85.67%). ¹H-NMR (400 MHz, Chloroform-d) δ 4.30-3.48(m, 5H), 2.75-2.56 (m, 1H), 2.24-1.90 (m, 3H), 1.86-1.65 (m, 1H), 1.47(s, 9H).

Step 4

A mixture of 57-D (720 mg, 2.74 mmol), phthalimide (402 mg, 2.73 mmol)and triphenylphosphine (1.61 g, 6.15 mmol) in THF (45 ml) was cooled inan ice bath. Diisopropyl azodicarboxylate, 95% (1.22 ml, 6.19 mmol), wasadded. The mixture was then warmed up to room temperature and stirredfor 20 h. After, the reaction mixture was concentrated and the residuedissolved in ether, cooled in an ice bath and stirred for 1.5 h. Afterthe solids were filtered off, the filtrate was concentrated. The residuewas purified by silica column chromatography (40-60% EtOAc/hexanes) toafford the phthalimide adduct (1.07 g, 99.7%). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₀H₂₂F₂N₂O₄: 392.4; found: 393.204.

A solution of the phthalimide adduct (1.07 g, 2.73 mmol) and hydrazinehydrate (0.54 mL, 11.15 mmol) in ethanol (10 ml) was stirred at 70° C.for 2 hours. The reaction mixture was cooled in an ice bath and ether(10 ml) was added. The mixture was stirred for 30 min. The solid formedwas filtered off and the filtrate was concentrated under vacuum todryness to afford crude 57-E.

Step 5

A mixture of crude 57-E (709 mg, 2.7 mmol) compound 38-F (Example 38)(936 mg, 2.7 mmol) and NaHCO₃ (454 mg, 5.41 mmol) in water (15 mL) andEtOH (15 mL) was stirred for 20 h. The reaction mixture was concentratedunder vacuum and the residue was partitioned between water and EtOAc.The aqueous layer was re-extracted with EtOAc and the combined organiclayers were dried (Na₂SO₄) and concentrated. The residue (1.5 g) wasdissolved in CH₂Cl₂ (7 mL) and 4N HCl in dioxane (26.9 mL) was added.After 1.5 hours the mixture was concentrated to dryness, co-evaporatedwith toluene and dried in vacuum. The crude residue (1.3 g) and DBU (2ml, 13.4 mmol) in toluene (25 ml) was stirred at 110° C. After 35minutes the mixture was concentrated and the residue was purified bysilica column chromatography (5-15% MeOH/EtOAc) to afford 57-F (426 mg,36.17%). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₃H₂₂F₂N₂O₅: 444.43;found: 445.280.

Step 6

The mixture of compound 57-F (426 mg, 0.96 mmol) in MeOH (7 ml) and THF(7 ml) was stirred at room temperature as 1M KOH (5.06 ml) was added.After 30 minutes the reaction mixture was neutralized with 1N HCl,extracted into EtOAc (2×) and the combined organic extracts were dried(Na₂SO₄) and concentrated to crude 57-G.

Step 7

A suspension of the crude residue 57-G (189 mg),2,4,6-trifluorobenzylamine (95 mg, 0.59 mmol) and HATU (276 mg, 0.73mmol) in dichloromethane (3 ml) was stirred at room temperature asN,N-diisopropylethylamine (DIPEA) (0.59 ml, 3.4 mmol) was added. After 1h the mixture was diluted with water, extracted into EtOAc (3×). Thecombined organic phases were dried (Na₂SO₄) and concentrated to 57-H.LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₈H₂₂F₅N₃O₄: 559.48; found:560.24.

Step 8

Compound 57-H (150 mg, 0.27 mmol) was dissolved in TFA (2 mL), stirredat room temperature for 10 min, and the solution was concentrated. Theresidue was purified by reverse phase HPLC (Gemini, 15 to 60%ACN/H₂O+0.1% TFA), to afford compound 57 (85 mg, 67.5%). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₁₆F₅N₃O₄: 469.36; found: 470.229.¹H-NMR (400 MHz, DMSO-d₆) δ 10.41 (t, J=5.6 Hz, 1H), 8.20 (s, 1H), 7.12(t, J=8.7 Hz, 2H), 4.79 (s, 1H), 4.48 (m, 3H), 4.10 (m, 2H), 3.02 (d,J=5.7 Hz, 1H), 2.33 (m, 1H), 2.22-1.97 (m, 2H), 1.85 (d, J=11.0 Hz, 1H),1.21 (s, 1H). ¹⁹F NMR (376 MHz, DMSO-d) δ −69.88, −71.77, −74.09, −88.33(dd, J=222.6, 23.8 Hz), −109.15˜−109.60 (m), −110.04, −112.44 (t, J=7.6Hz).

Example 58 Preparation of Compound 58(1R,4R,12aR)—N-(3-chloro-2,4-difluorobenzyl)-2,2-difluoro-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A suspension of the crude residue 57-G (120 mg),3-chloro,2,4-difluorobenzylamine (67 mg, 0.38 mmol) and HATU (175 mg,0.46 mmol) in dichloromethane (3 ml) was stirred at room temperature asN,N-diisopropylethylamine (DIPEA) (0.38 ml, 0.28 mmol) was added. After1 h the mixture was diluted with water, extracted into EtOAc (3×) thecombined organic phases were dried (Na₂SO₄) and concentrated to yield58-A. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₈H₂₂ClF₄N₃O₄: 575.94;found: 576.394.

Step 2

Compound 58-A (166 mg) was dissolved in TFA (2 mL), stirred at roomtemperature for 10 min, and the solution was concentrated. The residuewas purified by reverse phase HPLC (Gemini, 15 to 700/% ACN/H₂O+0.1%TFA), to afford compound 57 (60 mg, 42.8%). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₁H₁₆ClF₄N₃O₄: 485.82; found: 486.135. ¹H-NMR (400 MHz,DMSO-d6) δ 10.77 (t, J=6.0 Hz, 1H), 7.77 (s, 1H), 7.28 (m, 2H), 4.77 (s,1H), 4.64-4.40 (m, 2H), 4.27 (d, J=9.1 Hz, 1H), 3.93 (m, 2H), 2.95 (d,J=5.8 Hz, 1H), 2.51 (s, 1H), 2.42-2.17 (m, 1H), 2.14-1.89 (m, 2H), 1.77(m, 1H). ¹⁹F-NMR (376 MHz, DMSO-d₆) δ −87.63, −88.23, −108.67, −109.27,−116.42 (t, J=7.0 Hz), −118.48 (d, J=7.8 Hz).

Example 59 Preparation of Compound 59(1R,2R,4R,12aR)-2-fluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A solution of 57-B (1.9 g, 7.06 mmol) in methanol (35 mL) was stirred at0° C. as sodium borohydride (667 mg, 17.64 mmol) was added portionwiseand the resulting mixture was stirred at room temperature for 30 min.The reaction mixture was cooled in an ice bath, quenched by addition ofwater and concentrated. The residue was partitioned between water andEtOAc. The aqueous layer was re-extracted with EtOAc and the combinedorganic layers were dried (Na₂SO₄) and concentrated. The residue waspurified by silica column chromatography (30-60% EtOAc/hexanes) toafford 59-A (1.49 g). ¹H-NMR (400 MHz, chloroform-d) δ 4.57 (s, 1H),4.52-4.42 (m, 2H), 4.28 (s, 1H), 4.14 (s, 1H), 3.72 (d, J=2.1 Hz, 3H),2.74 (s, 1H), 2.08-1.87 (m, 2H), 1.43 (d, J=23.1 Hz, 10H) and 57-A (96mg): ¹H-NMR (400 MHz, chloroform-d) δ 4.65-4.40 (m, 2H), 4.34-4.02 (m,1H), 3.73 (d, J=2.3 Hz, 3H), 2.74 (t, J=5.3 Hz, 1H), 2.12-1.55 (m, 3H),1.52-1.18 (m, 11H).

Step 2

To a solution of 59-A (686 mg, 2.53 mmol) in N,N-dimethylformamide (5ml) was added tert-butylchlorodiphenylsilane (0.723 mL, 2.78 mmol) andimidazole (516 mg, 7.56 mmol). After 18 h, the mixture was diluted withwater, extracted into EtOAc (2×), and the organic fractions werecombined, dried (Na₂SO₄), and concentrated. The residue was purified bysilica column chromatography (10-20%, EtOAc/hexanes) to afford 59-C.LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₉H₃₉NO₅Si: 509.71; found:510.793.

Step 3

A solution of 59-C (1.23 g, 2.41 mmol) in THF (13 ml) was stirred in anice bath as 2.0 M LiBH₄ in THF (2.42 mL, 4.84 mmol)) was added and theresulting mixture was stirred at room temperature for 18 h. After thereaction mixture was diluted with ethyl acetate water was added slowly,two phases were separated, and the separated aqueous fraction wasextracted with ethyl acetate. The two organic fractions were washed withwater, combined, dried (Na₂SO₄), and concentrated. The residue waspurified by silica column chromatography (20-40% EtOAc/hexanes) toafford 59-D. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₅H₃₉NO₄Si: 481.7;found: 482.741.

Step 4

A mixture of 59-D (963 mg, 2.0 mmol), phthalimide (482 mg, 3.28 mmol)and triphenylphosphine (1.18 g, 4.49 mmol) in THF (50 ml) was cooled inan ice bath. Diisopropyl azodicarboxylate, 95% (0.89 mL, 4.52 mmol) wasadded. The mixture was then warmed up to room temperature and stirredfor 20 h. After, the reaction mixture was concentrated and the residuedissolved in ether, cooled in an ice bath and stirred for 1.5 h. After,the solids were filtered off and the filtrate was concentrated. Theresidue was purified by silica column chromatography (10-30%EtOAc/hexanes) to afford the phthalimide adduct. LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₃₆H₄₂N₂O₅Si: 610.81; found: 611.935.

A solution of the phthalimide adduct (1.2 g, 1.97 mmol) and hydrazinehydrate (0.4 ml, 8.03 mmol) in ethanol (12 ml) was stirred at 70° C. for2 h. The reaction mixture was cooled in an ice bath and ether (10 ml)was added, the mixture was stirred for 30 min. The solid formed wasfiltered off and the filtrate was concentrated under vacuum to drynessto afford 59-E. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₅H₄₀N₂O₃Si:480.71; found: 481.356.

Step 5

A mixture of crude 59-E (770 mg, 1.60 mmol), compound 38-F (Example 38)(555 mg, 1.60 mmol) and NaHCO₃ (269 mg, 3.20 mmol) in water (12 mL) andEtOH (12 mL) was stirred for 20 h. The reaction mixture was concentratedunder vacuum and the residue was partitioned between water and EtOAc.The aqueous layer was re-extracted with EtOAc and the combined organiclayers were dried (Na₂SO₄) and concentrated.

The residue (1.29 g) was dissolved in CH₂Cl₂ (4 mL) and 4N HCl indioxane (15.6 mL) was added. After 1.5 hours the mixture wasconcentrated to dryness, co-evaporated with toluene and dried in vacuum.LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₄₁H₄₈N₂O₇Si: 708.91; found:709.782.

The crude residue (1.09 mg) and DBU (1.17 ml, 7.8 mmol) in toluene (20ml) was stirred at 110° C. After 35 min the mixture was concentrated andthe residue was purified by silica column chromatography (5-15%MeOH/EtOAc) to afford 59-F. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₃₉H₄₂N₂O₆Si: 662.85; found: 663.677.

Step 6

A mixture of 59-F (680 mg, 1.03 mmol) in MeOH (10 ml) and THF (10 ml)was stirred at room temperature as 1M KOH (5.42 ml) was added. After 30min the reaction mixture was neutralized with 1N HCl, extracted intoEtOAc (2×) and the combined organic extracts were dried (Na₂SO₄) andconcentrated. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₃₇H₃₈N₂O₆Si:634.79; found: 635.466.

A suspension of the crude residue (650 mg), 2,4,6-trifluorobenzylamine(214 mg, 1.33 mmol) and HATU (623 mg, 1.64 mmol) in dichloromethane (6ml) was stirred at room temperature as N,N-diisopropylethylamine (DIPEA)(1.34 ml, 7.68 mmol) was added. After 2 h, the mixture was diluted withwater, extracted into EtOAc (3×) and the combined organic phases weredried (Na₂SO₄), concentrated and the residue was purified by silicacolumn chromatography (50-75% EtOAc/hexanes) to afford 59-G. LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₄₄H₄₂F₃N₃O₅Si: 777.9; found: 778.566.

Step 7

A solution of 59-G (648 mg, 0.83 mmol) in THF (10 ml) was stirred in anice bath as 1.0 M tetrabutylammonium fluoride in THF (0.83 ml) was addeddropwise and the resulting mixture was stirred at room temperature for30 min. Additional 1.0 M tetrabutylammonium fluoride in THF (0.1 ml) wasadded dropwise. After 30 minutes, the reaction mixture was concentratedunder vacuum and the residue was diluted with EtOAc, washed with water,dried (Na₂SO₄), concentrated and the residue was purified by silicacolumn chromatography (5% MeOH/EtOAc). A solution of the residue (290mg, 0.54 mmol) in dichloromethane (3 ml) was cooled to −78° C. asdiethylaminosulfur trifluoride (0.09 mL, 0.65 mmol) was added dropwiseand the reaction was warmed to room temperature and stirred for 2.5 h.The reaction was cooled in an ice bath, quenched with saturated NaHCO₃,two phases were separated, and the separated aqueous fraction wasextracted with CH₂Cl₂. The two organic fractions were combined, dried(Na₂SO₄), and concentrated. The residue was purified by silica columnchromatography (1% MeOH/EtOAc) to afford 59-H. LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₈H₂₃F₄N₃O₄: 541.49; found: 542.320.

Step 8

Compound 59-H (103 mg, 0.19 mmol) was dissolved in TFA (1.4 mL) at roomtemperature for 15 min, and the solution was concentrated. The residuewas suspended in DMF, filtered off, and the precipitated product waswashed with water, dried under vacuum to afford compound 59. LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₁₇F₄N₃O₄: 451.37; found: 452.226.¹H-NMR (400 MHz, DMSO-d6) δ 11.53 (s, 1H), 10.35 (t, J=5.8 Hz, 1H), 8.34(s, 1H), 7.18 (t, J=8.6 Hz, 2H), 5.15-4.88 (m, 1H), 4.73 (d, J=3.3 Hz,1H), 4.49 (m, 3H), 4.04 (t, J=12.4 Hz, 1H), 3.65 (dd, J=12.4, 3.7 Hz,1H), 2.95-2.76 (m, 1H), 2.26-2.03 (m, 1H), 1.96-1.64 (m, 3H). ¹⁹F-NMR(376 MHz, DMSO-d₆) δ −73.93, −74.74 (d, J=28.8 Hz), −109.31 (m), −112.51(m), −165.65 (m).

Example 60 Preparation of Compound 60(1R,4S,12aR)—N-(2,3-dichlorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

To a solution of dimethyl 3-methoxy-4-oxo-4H-pyran-2,5-dicarboxylate(5.5 g, 23 mmol) in MeOH (100 mL) was added 41-E (Example 41) (5 g, 22mmol) and sodium bicarbonate (3.6 g, 43 mmol). The solution was stirredat room temperature for 1.5 h. 4M HCl (in dioxane, 55 mL, 221 mmol) wasadded and the solution was heated to 50° C. for 2 h. The reaction wascooled to room temperature and concentrated in vacuo. The resulting oilwas dissolved in sodium bicarbonate and washed with EtOAc. The aqueouslayers were then extracted with CH₂Cl₂ (4×). The combined CH₂Cl₂extractions were dried over Na₂SO₄ and concentrated to provide 60-A.LCMS-ESI (m/z): [M+H]⁺ calculated for C₁₆H₁₉N₂O₅: 319.13; found: 319.20.

Step 2

To a suspension of 60-A (3.7 g, 11.6 mmol) in MeOH (12 mL) and THF (23mL) was added aqueous KOH (2M, 15.7 mL, 31.4 mmol). The resultingsolution was stirred at room temperature for 10 min. Volatiles wereremoved in vacuo, and the resulting aqueous layer was acidified with 1NHCl. The resulting white solid was filtered, washed with water, anddried in vacuo to provide 60-B. ¹H-NMR (400 MHz, Chloroform-d) δ 8.36(s, 1H), 5.01 (d, J=2.7 Hz, 1H), 4.12 (s, 4H), 3.90 (t, J=12.2 Hz, 1H),3.78 (dd, J=12.1, 3.1 Hz, 1H), 2.69 (s, 1H), 1.95-1.71 (m, 4H),1.70-1.54 (m, 2H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₅H₁₇N₂O₅:305.11; found: 305.15.

Step 3

To a solution of 60-B (0.10 g, 0.33 mmol) in CH₂Cl₂ (3.5 mL) was added(2,3-dichlorophenyl)methanamine (0.12 g, 0.70 mmol), HATU (0.25 g, 0.66mmol), and N,N-diisopropylethylamine (DIPEA) (0.29 mL, 1.64 mmol). Theresulting solution was stirred at room temperature until judged completeby LC/MS. The reaction mixture was diluted with CH₂Cl₂ and washed with1N HCl. The aqueous layer was back-extracted with CH₂Cl₂, and thecombined organic layers were dried over Na₂SO₄ and concentrated invacuo. The crude material was dissolved in hot DMF and allowed toprecipitate upon cooling. Filtration provided 60-C. LCMS-ESI⁺ (m/m):[M+H]⁺ calculated for C₂₂H₂₂Cl₂N₃O₄: 462.10; found: 462.14.

Step 4

To a slurry of 60-C (0.11 g, 0.24 mmol), in acetonitrile (4.5 mL), wasadded magnesium bromide (0.089 g, 0.48 mmol). The reaction mixture washeated to 45° C. for 2.5 h and then cooled to room temperature. Theslurry was diluted with CH₂Cl₂ and washed with 1N HCl and brine. Theaqueous layers were back-extracted with CH₂Cl₂ (2×) and the combinedorganic layers were dried over Na₂SO₄ and concentrated in vacuo. Thecrude solid was triturated with methanol and filtered to provide 60.¹H-NMR (400 MHz, DMSO-d6) δ 11.72 (s, 1H), 10.50 (t, 1H), 8.34 (s, 1H),7.55 (dd, 1H), 7.40-7.24 (m, 2H), 4.67 (s, 1H), 4.61 (d, 2H), 4.45 (dd,1H), 3.95 (t, 1H), 3.84-3.73 (m, 1H), 1.86-1.67 (m, 3H), 1.66-1.40 (m,4H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀Cl₂N₃O₄: 448.08;found: 448.18.

Example 61 Preparation of Compound 61(1R,4S,12aS)—N-(3-chloro-2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

61 was prepared analogously to Example 60, substituting(1S,3S,4R)-tert-butyl3-(aminomethyl)-2-azabicyclo[2.2.1]heptane-2-carboxylate (prepared inExample 55) for 41-E, and (3-chloro-2,4-difluorophenyl)methanamine for(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, DMSO-d6) δ 11.85 (s,1H), 10.45 (t, 1H), 8.40 (s, 1H), 7.37 (td, 1H), 7.27 (td, 1H),4.63-4.46 (m, 4H), 4.17 (t, 1H), 4.04 (dt, 1H), 1.76 (d, 1H), 1.73-1.54(m, 5H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₉ClF₂N₃O₄: 450.10;found: 450.15.

Example 62 Preparation of Compound 62′(2R,5S,13aR)—N-(4-fluoro-2-(trifluoromethyl)benzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 62 was prepared in a similar manner to compound 42 using(4-fluoro-2-(trifluoromethyl)phenyl)methanamine in place of(2,4,6-trifulorophenylphenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d)δ 10.50 (s, 1H), 8.38 (s, 1H), 7.57 (dd, 1H), 7.36 (dd, 1H), 7.19 (td,1H), 5.40-5.28 (m, 2H), 4.79 (t, 2H), 4.69 (s, 1H), 4.25 (dd, 1H), 4.03(dd, 1H), 2.17-1.98 (m, 4H), 1.96-1.84 (m, 1H), 1.61 (dt, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₂H₂₀F₄N₃O₅: 482.13; found: 482.145.

Example 63 Preparation of Compound 63(2R,5S,13aR)—N-(2-chloro-4-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 63 was prepared in a similar manner to compound 42 using(2-chloro-4-fluorophenyl)methanamine in place of(2,4,6-trifulorophenylphenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d)δ 10.48 (s, 1H), 8.45 (s, 1H), 7.39 (dd, 1H), 7.12 (dd, 1H), 6.93 (td,1H), 5.37 (d, 1H), 5.31 (t, 1H), 4.68 (s, 3H), 4.29 (d, 1H), 4.04 (t,1H), 2.21-2.01 (m, 4H), 1.97-1.82 (m, 1H), 1.67-1.56 (m, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₂₀ClFN₃O₅: 448.10; found: 448.143.

Example 64 Preparation of Compound 64(2R,5S,13aR)-8-hydroxy-7,9-dioxo-N-(2,4,5-trifluorobenzyl)-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 64 was prepared in a similar manner to compound 42 using(2,4,5-trifluorophenyl)methanamine in place of(2,4,6-trifulorophenylphenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d)δ 10.42 (s, 1H), 8.42 (s, 1H), 7.19 (ddd, 1H), 6.91 (td, 1H), 5.38 (dd,1H), 5.31 (t, 1H), 4.69 (s, 1H), 4.61 (d, 2H), 4.29 (dd, 1H), 4.05 (dd,1H), 2.18-2.02 (m, 4H), 1.96-1.84 (m, 1H), 1.66-1.56 (m, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₁₉F₃N₃O₅: 450.12; found: 450.119.

Example 65 Preparation of Compound 65(2R,5S,13aR)—N-(5-chloro-2,4-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 65 was prepared in a similar manner to compound 42 using(5-chloro-2,4-difluorophenyl)methanamine in place of(2,4,6-trifulorophenylphenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d)δ 10.47 (t, 1H), 8.41 (s, 1H), 7.40 (dd, 1H), 6.90 (t, 1H), 5.37 (dd,1H), 5.31 (t, 1H), 4.69 (s, 1H), 4.62 (d, 2H), 4.28 (d, 1H), 4.04 (dd,1H), 2.17-2.02 (m, 4H), 1.94-1.86 (m, 1H), 1.61 (dt, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₁₉ClF₂N₃O₅: 466.09; found: 466.107.

Example 66 Preparation of Compound 66(1R,4S,12aR)—N-(3,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 66 was prepared in a similar manner to compound 60 using(3,4-difluorophenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 10.59(s, 1H), 7.24-7.16 (m, 2H), 7.14-7.04 (m, 2H), 4.91 (s, 1H), 4.58 (d,3H), 3.94-3.82 (m, 1H), 3.79 (d, 1H), 1.99-1.81 (m, 4H), 1.76 (d, 1H),1.70-1.60 (m, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₀F₂N₃O₄:416.13; found: 416.415.

Example 67 Preparation of Compound 67(1R,4S,12aR)—N-(4-fluoro-2-(trifluoromethyl)benzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 67 was prepared in a similar manner to compound 60 using(4-fluoro-2-(trifluoromethyl)phenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 11.72(s, 1H), 10.55 (s, 1H), 8.29 (s, 1H), 7.61 (s, 1H), 7.36 (dd, 1H), 7.18(td, 1H), 4.91 (s, 1H), 4.80 (d, 3H), 4.11 (s, 1H), 1.99-1.80 (m, 4H),1.76 (d, 1H), 1.71-1.47 (m, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₀F₄N₃O₄: 466.13; found: 466.297.

Example 68 Preparation of Compound 68(1R,4S,12aR)—N-(2-chloro-4-fluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 68 was prepared in a similar manner to compound 60 using(2-chloro-4-fluorophenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 11.68(s, 1H), 10.52 (s, 1H), 8.27 (s, 1H), 7.44-7.37 (m, 1H), 7.11 (dd, 1H),6.93 (td, 1H), 4.90 (s, 1H), 4.68 (d, 2H), 4.16-4.01 (m, 1H), 3.88-3.70(m, 2H), 2.00-1.79 (m, 4H), 1.75 (d, 1H), 1.70-1.57 (m, 2H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₂₀ClFN₃O₄: 432.10; found: 432.214.

Example 69 Preparation of Compound 69(1R,4S,12aR)—N-(3-chloro-2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 69 was prepared in a similar manner to compound 60 using(3-chloro-2,4-difluorophenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 11.71(s, 1H), 10.48 (s, 1H), 8.26 (s, 1H), 7.27 (s, 1H), 6.92 (td, 1H), 4.90(s, 1H), 4.66 (d, 2H), 4.08 (s, 1H), 3.91-3.69 (m, 2H), 2.01-1.79 (m,3H), 1.75 (d, 1H), 1.71-1.44 (m, 2H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₁H₁₉ClF₂N₃O₄: 450.10; found: 450.27.

Example 70 Preparation of Compound 70(1R,4S,12aR)—N-(2-fluoro-3-methylbenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 70 was prepared in a similar manner to compound 60 using(2-fluoro-3-methylphenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 11.62(s, 1H), 10.39 (s, 1H), 8.30 (s, 1H), 7.19 (t, 1H), 7.07 (t, 1H), 6.96(t, 1H), 4.89 (d, 1H), 4.67 (d, 2H), 4.08 (s, 1H), 3.88-3.67 (m, 2H),2.26 (d, 3H), 1.97-1.79 (m, 3H), 1.78-1.39 (m, 3H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₂H₂₃FN₃O₄: 412.16; found: 412.26.

Example 71 Preparation of Compound 71(1R,4S,12aR)—N-(3,6-dichloro-2-fluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 71 was prepared in a similar manner to compound 60 using(3,6-dichloro-2-fluorophenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 11.62(s, 1H), 10.47 (t, 1H), 8.29 (s, 1H), 7.13 (dd, 1H), 4.88 (s, 1H),4.85-4.73 (m, 2H), 4.09 (d, 1H), 3.88-3.68 (m, 2H), 1.99-1.53 (m, 8H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₉Cl₂FN₃O₄: 466.07; found:466.257.

Example 72 Preparation of Compound 72(1R,4S,12aR)—N-(3-chlorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 72 was prepared in a similar manner to compound 60 using(3-chlorophenyl)methanamine in place of (2,3-dichlorophenyl)methanamine.¹H-NMR (400 MHz, DMSO-d6) δ 11.75 (s, 1H), 10.44 (t, 1H), 8.38 (s, 1H),7.42-7.22 (m, 4H), 4.68 (s, 1H), 4.54 (d, 2H), 4.48 (dd, 1H), 3.97 (t,1H), 3.81 (dd, 1H), 2.58 (s, 1H), 1.87-1.69 (m, 3H), 1.68-1.51 (m, 2H),1.46 (d, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₂₁ClN₃O₄:414.11; found: 414.21.

Example 73 Preparation of Compound 73(1R,4S,12aR)—N-(3-chloro-2,6-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 73 was prepared in a similar manner to compound 60 using(3-chloro-2,6-difluorophenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, DMSO-d6) δ 11.71 (s,1H), 10.46 (t, 1H), 8.34 (s, 1H), 7.60 (td, 1H), 7.19 (td, 1H), 4.67 (s,1H), 4.62 (d, 2H), 4.44 (dd, 1H), 3.95 (t, 1H), 3.78 (dd, 1H), 2.57 (s,1H), 1.86-1.68 (m, 3H), 1.67-1.49 (m, 2H), 1.45 (d, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₁₉ClF₂N₃O₄: 450.10; found: 450.16.

Example 74 Preparation of Compound 74(1R,4S,12aR)—N-(2-fluoro-3-(trifluoromethyl)benzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 74 was prepared in a similar manner to compound 60 using(2-fluoro-3-(trifluoromethyl)phenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, DMSO-d6) δ 11.76 (s,1H), 10.48 (t, 1H), 8.36 (s, 1H), 7.68 (q, 2H), 7.38 (t, 1H), 4.68 (s,1H), 4.65 (d, 2H), 4.47 (dd, 1H), 3.96 (t, 1H), 3.80 (dd, 1H), 2.57 (s,1H), 1.88-1.69 (m, 3H), 1.67-1.50 (m, 2H), 1.45 (d, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₂H₂₀F₄N₃O₄: 466.13; found: 466.142.

Example 75 Preparation of Compound 75(1R,4S,12aR)—N-(3-chloro-4-fluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 75 was prepared in a similar manner to compound 60 using(3-chloro-4-fluorophenyl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, DMSO-d6) δ 11.75 (s,1H), 10.43 (t, 1H), 8.38 (s, 1H), 7.51 (dd, 1H), 7.42-7.28 (m, 2H), 4.68(s, 1H), 4.51 (d, 2H), 4.47 (dd, 1H), 3.97 (t, 1H), 3.80 (dd, 1H), 2.58(s, 1H), 1.86-1.68 (m, 3H), 1.68-1.52 (m, 2H), 1.46 (d, 1H). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₂₁H₂₀ClFN₃O₄: 432.10; found: 432.159.

Example 76 Preparation of Compound 76(1R,4S,12aR)—N-((3,5-difluoropyridin-2-yl)methyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 76 was prepared in a similar manner to compound 60 using(3,5-difluoropyridin-2-yl)methanamine in place of(2,3-dichlorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 10.80(s, 1H), 8.81 (s, 1H), 8.33 (d, 1H), 7.20 (td, 1H), 4.90 (s, 1H), 4.82(s, 2H), 4.28 (d, 1H), 3.92-3.75 (m, 2H), 3.48 (s, 2H), 1.98-1.80 (m,3H), 1.77 (d, 1H), 1.71-1.58 (m, 2H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₀H₁₉F₂N₄O₄: 417.13; found: 417.189.

Example 77 Preparation of Compound 77(1R,4S,12aR)-7-hydroxy-6,8-dioxo-N—((R)-1-(2,4,6-trifluorophenyl)ethyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1,2′-d]pyrazine-9-carboxamide

Step 1

A 50-mL round bottom flask was charged with 77-A (0.15 g, 0.39 mmol),(R)-1-(2,4,6-trifluorophenyl)ethanamine (0.14 g, 0.78 mmol),N,N-diisopropylethylamine (DIPEA) (0.25 g, 1.97 mmol) and HATU (0.29 g,0.79 mmol) in DCM (10 ml). The reaction mixture was stirred at roomtemperature for 1 h. The reaction mixture was concentrated down,re-dissolved in EtOAc (50 mL), washed with saturated NaHCO₃ (2×),saturated NH₄Cl and dried over Na₂SO₄. After concentration, the crudewas purified by column chromatography on silica gel with hexane-EtOAc toobtain 77-B as a white solid. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 538.

Step 2

A 50-mL round bottom flask was charged with 77-B (0.20 g, 0.37 mmol) inTFA (2 mL). The reaction mixture was stirred at room temperature for 30min. The solution was concentrated and the residue was purified by flashchromatography using EtOAc-20% MeOH in EtOAc as eluents to affordcompound 77. ¹H-NMR (400 MHz, Chloroform-d) δ 10.67 (d, J=8.2 Hz, 1H),8.22 (s, 1H), 6.61 (t, J=8.4 Hz, 2H), 5.60 (dd, J=8.1, 6.9 Hz, 1H), 4.85(s, 1H), 3.82 (t, J=12.2 Hz, 1H), 3.71 (dd, J=12.4, 3.4 Hz, 1H),2.75-2.55 (m, 3H), 1.97-1.57 (m, 9H). ¹⁹F-NMR (376 MHz, Chloroform-d)6-109.65˜−111.29 (m), −111.76˜−113.09 (m). LCMS-ESI⁺ (m/z): [M+H]⁺found: 448.

Example 78 Preparation of Compound 78(2R,13aR)-8-hydroxy-7,9-dioxo-N—((R)-1-(2,4,6-trifluorophenyl)ethyl)-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Step 1

A 50-mL round bottom flask was charged with 78-A (0.30 g, 0.94 mmol),(R)-1-(2,4,6-trifluorophenyl) ethanamine (0.39 g, 1.87 mmol),N,N-diisopropylethylamine (DIPEA) (0.61 g, 4.87 mmol) and HATU (0.71 g,1.87 mmol) in DCM (10 ml). The reaction mixture was stirred at roomtemperature for 1 hour. The reaction mixture was concentrated down,re-dissolved in EtOAc (50 mL), washed with saturated NaHCO₃ (2×),saturated NH₄Cl and dried over Na₂SO₄. After concentration, the crudewas purified by column chromatography on silica gel with hexane-EtOAc toobtain 78-B as a white solid. LCMS-ESI⁺ (m/z): [M+H]⁺; found: 478.

Step 2

A 50-mL round bottom flask was charged with 78-B (0.4 g, 0.84 mmol) andmagnesium bromide (0.4 g, 2.2 mmol) in acetonitrile (5 mL). The reactionmixture was heated to 50° C. After 10 minutes, the reaction mixture wascooled to 0° C. and 1 N hydrochloric acid (4 mL) was added in. Morewater (˜5 mL) was added and the solid was filtrated and washed withwater and dried to afford compound 78. ¹H-NMR (400 MHz, Chloroform-d) δ12.30 (s, 1H), 10.59 (d, J=8.3 Hz, 1H), 8.21 (s, 1H), 6.60 (t, J=8.4 Hz,2H), 5.59 (t, J=7.4 Hz, 1H), 5.37 (dd, J=9.4, 4.1 Hz, 1H), 5.31-5.09 (m,1H), 4.64 (t, J=3.0 Hz, 1H), 4.20 (dd, J=12.9, 4.1 Hz, 2H), 3.96 (dd,J=12.8, 9.4 Hz, 2H), 2.21-1.85 (m, 4H), 1.71-1.43 (m, 3H). ¹⁹F-NMR (376MHz, Chloroform-d) δ −110.37 (tt, J=8.7, 6.1 Hz), −112.19 (t, J=7.2 Hz).LCMS-ESI⁺ (m/z): [M+H]⁺ found: 464.

Example 79 Preparation of Compound 79(1R,4S,12aR)-7-hydroxy-6,8-dioxo-N-(2,4,5-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 50-mL round bottom flask was charged with 79-A (0.12 g, 0.32 mmol),(2,4,5-trifluorophenyl)methanamine (0.10 g, 0.63 mmol),N,N-diisopropylethylamine (DIPEA) (0.20 g, 1.58 mmol) and HATU (0.24 g,0.63 mmol) in DCM (10 ml). The reaction mixture was stirred at roomtemperature for 1 h. The reaction mixture was concentrated down,re-dissolved in EtOAc (50 mL), washed with saturated NaHCO₃ (2×),saturated NH₄Cl and dried over Na₂SO₄. After concentration, the crudewas purified by column chromatography on silica gel with hexane-EtOAc toobtain 79-B as a white solid. LCMS-ESI⁺ (m/z): [M+H]⁺; found: 524.

Step 2

A 50-mL round bottom flask was charged with 79-B (0.15 g, 0.29 mmol) inTFA (2 mL). The reaction mixture was stirred at room temperature for 30min. The solution was concentrated and the residue was purified by flashchromatography using EtOAc-20% MeOH in EtOAc as eluents to affordcompound 79. ¹H-NMR (400 MHz, Chloroform-d) δ 11.70 (s, 1H), 10.65-10.18(m, 1H), 8.27 (s, 1H), 7.26 (m, 1H), 6.90 (td, J=9.7, 6.4 Hz, 1H), 4.89(s, 1H), 4.60 (d, J=6.0 Hz, 2H), 4.09 (dd, J=11.4, 2.6 Hz, 1H),3.96-3.66 (m, 2H), 2.68 (s, 1H), 2.15-1.43 (m, 6H). ¹⁹F-NMR (376 MHz,Chloroform-d) δ 120.53˜−120.85 (m), −134.68˜−136.79 (m), −142.26˜−144.11(m). LCMS-ESI⁺ (m/z): [M+H]⁺ found: 434.

Example 80 Preparation of Compound 80(1R,4S,12aR)—N-(5-chloro-2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 50-mL round bottom flask was charged with 80-A (0.12 g, 0.32 mmol),(5-chloro-2,4-difluorophenyl)methanamine (0.11 g, 0.63 mmol),N,N-diisopropylethylamine (DIPEA) (0.20 g, 1.58 mmol) and HATU (0.24 g,0.63 mmol) in DCM (10 ml). The reaction mixture was stirred at roomtemperature for 1 h. The reaction mixture was concentrated down,re-dissolved in EtOAc (50 mL), washed with saturated NaHCO₃ (2×),saturated NH₄Cl and dried over Na₂SO₄. After concentration, the crudewas purified by column chromatography on silica gel with hexane-EtOAc toobtain 80-B as a white solid. LCMS-ESI⁺ (m/z): [M+H]⁺; found: 541.

Step 2

A 50-mL round bottom flask was charged with 80-B (0.14 g, 0.26 mmol) inTFA (2 mL). The reaction mixture was stirred at room temperature for 30minutes. The solution was concentrated and the residue was purified byflash chromatography using EtOAc-20% MeOH in EtOAc as eluents to affordcompound 80. ¹H-NMR (400 MHz, Chloroform-d) δ 10.46 (s, 1H), 8.27 (s,1H), 7.40 (t, J=7.8 Hz, 1H), 6.89 (t, J=9.1 Hz, 1H), 4.90 (s, 1H),4.78-4.48 (m, 2H), 4.08 (dd, J=11.3, 2.5 Hz, 1H), 3.95-3.63 (m, 2H),2.68 (s, 1H), 2.22-1.51 (m, 7H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ−113.37 (q, J=8.1 Hz), −116.37 (q, J=8.0 Hz). LCMS-ESI⁺ (m/z): [M+H]⁺found: 451.

Example 81 Preparation of Compound 81(1R,3S,4S,12aS)-3-fluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL round bottom flask was charged with 81-A (1.0 g, 3.7 mmol) inDCM (10 mL). The reaction mixture was cooled to 0° C. Diethylaminosulfurtrifluoride (DAST) (0.58 mL, 4.1 mmol) was slowly added in. Then thereaction mixture was stirred at room temperature for one hour. Themixture was cooled back to 0° C. Saturated NaHCO₃ (5 mL) was addeddropwise to quench the reaction. Then the reaction mixture was dilutedwith EtOAc (100 mL), washed with sat. NaHCO₃, brine, and dried overNa₂SO₄. After concentration, the residue was purified by flashchromatography using hexanes-EtOAc as eluents to afford 81-B. LCMS-ESI⁺(m/z): [M+H]⁺ found: 274.

Step 2

A 100-mL round bottom flask was charged with 81-B (0.8 g, 3.0 mmol) inTHF (10 mL). The reaction mixture was stirred at −78° C. 2.0 M LiBH₄ inTHF (3.2 mL, 6.4 mmol) was slowly added in. Then the reaction mixturewas warmed up and stirred at room temperature for 3 hours. Then thereaction mixture was diluted with EtOAc (100 mL) and treated slowly withwater (H₂ evolution). After the two phases were separated, the aqueousfraction was extracted with EtOAc and the two organic fractions werecombined, washed with water, and dried over Na₂SO₄. After concentration,the residue was purified by flash chromatography using hexanes-EtOAc aseluents to afford 81-C. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 246.

Step 3

A 100-mL round bottom flask was charged with 81-C (0.57 g, 2.3 mmol),triphenylphosphine (1.3 g, 5.1 mmol) and phthalimide (0.55 g, 3.7 mmol)in THF (15 mL). Then the reaction mixture was cooled to 0° C. withstirring. Diisopropyl azodicarboxylate (DIAD) (1.0 mL, 5.1 mmol) wasslowly added to the reaction mixture. The reaction mixture was stirredat room temperature for overnight. After concentration, the residue waspurified by flash chromatography using hexanes-EtOAc as eluents toafford 81-D. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 375.

Step 4

To a solution of 81-D (0.8 g, 2.1 mmol) EtOH (40 mL) was added hydrazinemonohydrate (0.6 mL). The reaction mixture was heated to 70° C. withstirring for 3 hours. After filtration to remove the solid, the filtratewas concentrated to afford 81-E. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 245.

Step 5

A 100-mL round bottom flask was charged with 81-E (0.49 g, 2.0 mmol) and81-F (0.7 g, 2.0 mmol) in ethanol (7 mL). Sodium bicarbonate (0.34 g,4.0 mmol) in water (7 mL) was added to the reaction mixture. Then thereaction mixture was stirred at room temperature for overnight. Themixture was diluted with EtOAc (50 mL) and washed with water (2×). Theaqueous fractions were extracted with EtOAc (1×), and the organicfractions were combined, dried (Na₂SO₄), and concentrated. The crude81-G was used for next step without further purification. LCMS-ESI⁺(m/z): [M+H]⁺ found: 573.

Step 6

A 100-mL round bottom flask was charged with 81-G (1.1 g, 1.9 mmol) in 4N HCl/dioxane (11 mL). Then the reaction mixture was stirred at roomtemperature for 1 hour. After concentration, 1.0 g intermediate wasobtained. The intermediate and DBU (1.3 g, 8.8 mmol) were dissolved intoluene (10 mL). The reaction mixture was heated to 110° C. withstirring for 1 hour. After concentration, the residue was purified byflash chromatography using hexanes-EtOAc as eluents to afford 81-H.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 413.

Step 7

A 100-mL round bottom flask was charged with 81-H (0.56 g, 1.4 mmol) inTHF (5 mL) and MeOH (5 mL). 1 N KOH (4 mL) was added to the reactionmixture. Then the reaction mixture was stirred at room temperature for 1hour. The reaction mixture was acidified by adding 1 N HCl (4 mL). Afterconcentration, the residue was co-evaporated with toluene (3×). Half ofthe crude acid, 2,4,6-trifluobenzylamine (0.2 g, 1.3 mmol),N,N-diisopropylethylamine (DIPEA) (0.41 g, 3.1 mmol) and HATU (0.48 g,1.25 mmol) were dissolved in DMF (10 mL). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted withEtOAc (100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl(2×) and dried over Na₂SO₄. After concentration, the crude was purifiedby column chromatography on silica gel with hexane-EtOAc to afford 81-I.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 542.

Step 8

A 50-mL round bottom flask was charged with 81-I (0.31 g, 0.58 mmol) inTFA (3 mL). The reaction mixture was stirred at room temperature for 30minutes. After concentration, the crude was purified by columnchromatography on silica gel with EtOAc-MeOH to afford compound 81.¹H-NMR (400 MHz, Chloroform-d) δ 10.29 (s, 1H), 8.31 (s, 1H), 6.65 (dd,J=8.7, 7.5 Hz, 2H), 5.05-4.75 (m, 2H), 4.65 (d, J=5.6 Hz, 2H), 4.11 (d,J=12.2 Hz, 1H), 3.83 (t, J=12.3 Hz, 1H), 3.56 (dd, J=12.3, 3.3 Hz, 1H),2.77 (s, 1H), 2.25-1.97 (m, 2H), 1.95 (d, J=11.0 Hz, 2H), 1.77 (d,J=11.2 Hz, 1H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −108.98 (t, J=8.2 Hz),−112.03 (t, J=7.2 Hz), −168.00. LCMS-ESI⁺ (m/z): found: 452.

Example 82 Preparation of Compound 82(1S,3R,4R,12aR)-3-fluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL round bottom flask was charged with 82-A (0.6 g, 2.1 mmol) inDCM (6 mL). The reaction mixture was cooled to 0° C. DAST (0.35 mL, 3.0mmol) was slowly added in. Then the reaction mixture was stirred at roomtemperature for one hour. The mixture was cooled back to 0° C. SaturatedNaHCO₃ (5 mL) was added drop wise to quench the reaction. Then thereaction mixture was diluted with EtOAc (100 mL), washed with sat.NaHCO₃, brine, and dried over Na₂SO₄. After concentration, the residuewas purified by flash chromatography using hexanes-EtOAc as eluents toafford 82-B. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 274.

Step 2

A 100-mL round bottom flask was charged with 82-B (0.4 g, 1.5 mmol) inTHF (10 mL). The reaction mixture was stirred at −78° C. 2.0 M LiBH₄ inTHF (1.6 mL, 3.2 mmol) was slowly added in. Then the reaction mixturewas warmed up and stirred at room temperature for 3 hours. Then thereaction mixture was diluted with EtOAc (100 mL) and added water slowly(H₂ evolution). After the two phases were separated, the aqueousfraction was extracted with EtOAc and the two organic fractions werecombined, washed with water and dried over Na₂SO₄. After concentration,the residue was purified by flash chromatography using hexanes-EtOAc aseluents to afford 82-C. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 246.

Step 3

A 100-mL round bottom flask was charged with 82-C (0.25 g, 1.0 mmol),triphenylphosphine (0.59 g, 2.2 mmol) and phthalimide (0.24 g, 1.6 mmol)in THF (10 mL). Then the reaction mixture was cooled to 0° C. withstirring. DIAD (0.44 mL, 2.2 mmol) was slowly added to the reactionmixture. The reaction mixture was stirred at room temperature forovernight. After concentration, the residue was purified by flashchromatography using hexanes-EtOAc as eluents to afford 82-D. LCMS-ESI⁺(m/z): [M+H]⁺ found: 375.

Step 4

To a solution of 82-D (0.35 g, 0.9 mmol) EtOH (20 mL) was addedhydrazine monohydrate (0.3 mL). The reaction mixture was heated to 70°C. with stirring for 3 hours. After filtration to remove the solid, thefiltrate was concentrated to afford 82-E. LCMS-ESI⁺ (m/z): [M+H]⁺ found:245.

Step 5

A 100-mL round bottom flask was charged with 82-E (0.21 g, 0.87 mmol)and 82-F (0.3 g, 0.87 mmol) in ethanol (7 mL). Sodium bicarbonate (0.15g, 1.7 mmol) in water (7 mL) was added to the reaction mixture. Then thereaction mixture was stirred at room temperature for overnight. Themixture was diluted with EtOAc (50 mL) and washed with water (2×). Theaqueous fractions were extracted with EtOAc, and the organic fractionswere combined, dried (Na₂SO₄), and concentrated. The crude 82-G was usedfor next step without further purification. LCMS-ESI⁺ (m/z): [M+H]⁺found: 573.

Step 6

A 100-mL round bottom flask was charged with 82-G (0.49 g, 0.86 mmol) in4 N HCl/dioxane (5 mL). Then the reaction mixture was stirred at roomtemperature for 1 hour. After concentration, 0.4 g intermediate wasobtained. The intermediate and DBU (0.6 g, 4.0 mmol) were dissolved intoluene (10 mL). The reaction mixture was heated to 110° C. withstirring for 1 hour. After concentration, the residue was purified byflash chromatography using hexanes-EtOAc as eluents to afford 82-H.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 413.

Step 7

A 100-mL round bottom flask was charged with 82-H (0.2 g, 0.49 mmol) inTHF (5 mL) and MeOH (5 mL). 1 N KOH (1.5 mL) was added to the reactionmixture. Then the reaction mixture was stirred at room temperature for 1hour. The reaction mixture was acidified by adding 1 N HCl (1.5 mL).After concentration, the residue was co-evaporated with toluene (3×).The crude acid, 2,4,6-trifluobenzylamine (0.15 g, 0.95 mmol),N,N-diisopropylethylamine (DIPEA) (0.31 g, 2.4 mmol) and HATU (0.36 g,0.95 mmol) were dissolved in DCM (10 mL). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted withEtOAc (100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl(2×) and dried over Na₂SO₄. After concentration, the crude was purifiedby column chromatography on silica gel with hexane-EtOAc to afford 82-I.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 542.

Step 8

A 50-mL round bottom flask was charged with 82-I (0.22 g, 0.41 mmol) inTFA (3 mL). The reaction mixture was stirred at room temperature for 30minutes. After concentration, the crude was purified by columnchromatography on silica gel with EtOAc-MeOH to afford compound 82.¹H-NMR (400 MHz, Chloroform-d) δ 10.25 (s, 1H), 8.28 (s, 1H), 6.65 (s,2H), 5.15-4.77 (m, 2H), 4.65 (s, 2H), 4.32-3.41 (m, 2H), 2.78 (s, 1H),1.86 (dd, J=144.8, 72.3 Hz, 6H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ−108.98 (t, J=8.2 Hz), −112.03 (t, J=7.2 Hz), −168.00. LCMS-ESI⁺ (m/z):found: 452.

Example 83 Preparation of Compound 83(1S,4R,12aS)-3,3-difluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL round bottom flask was charged with 83-A (1.0 g, 3.7 mmol) inDCM (20 mL). The reaction mixture was cooled to 0° C. Dess-Martinperiodinane (1.8 g, 4.2 mmol) was slowly added in. Then the reactionmixture was stirred at room temperature for 3 hours. Afterconcentration, the residue was purified by flash chromatography usinghexanes-EtOAc as eluents to afford 83-B. LCMS-ESI⁺ (m/z): [M+H]⁺ found:270.

Step 2

A 100-mL round bottom flask was charged with 83-B (0.85 g, 3.2 mmol) inDCM (15 mL). The reaction mixture was cooled to 0° C. DAST (1.5 mL, 11.3mmol) was slowly added in. Then the reaction mixture was stirred at roomtemperature overnight. The mixture was cooled back to 0° C. SaturatedNaHCO₃ (5 mL) was added dropwise to quench the reaction. Then thereaction mixture was diluted with EtOAc (100 mL), washed with sat.NaHCO₃, brine, and dried over Na₂SO₄. After concentration, the residuewas purified by flash chromatography using hexanes-EtOAc as eluents toafford 83-C. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 292.

Step 3

A 100-mL round bottom flask was charged with 83-C (0.44 g, 1.5 mmol) inTHF (6 mL). The reaction mixture was stirred at −78° C. 2.0 M LiBH₄ inTHF (1.6 mL, 3.2 mmol) was slowly added in. Then the reaction mixturewas warmed up and stirred at room temperature for 3 hours. Then thereaction mixture was diluted with EtOAc (100 mL) and added water slowly(H₂ evolution). After the two phases were separated, the aqueousfraction was extracted with EtOAc and the two organic fractions werecombined, washed with water, and dried over Na₂SO₄. After concentration,the residue was purified by flash chromatography using hexanes-EtOAc aseluents to afford 83-D. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 264.

Step 4

A 100-mL round bottom flask was charged with 83-D (0.17 g, 0.65 mmol),triphenylphosphine (0.37 g, 1.4 mmol) and phthalimide (0.15 g, 1.0 mmol)in THF (10 mL). Then the reaction mixture was cooled to 0° C. withstirring. DIAD (0.28 mL, 1.4 mmol) was slowly added to the reactionmixture. The reaction mixture was stirred at room temperature forovernight. After concentration, the residue was purified by flashchromatography using hexanes-EtOAc as eluents to afford 83-E. LCMS-ESI⁺(m/z): [M+H]⁺ found: 393.

Step 5

To a solution of 83-E (0.25 g, 0.64 mmol) EtOH (20 mL) was addedhydrazine monohydrate (0.3 mL). The reaction mixture was heated to 70°C. with stirring for 3 hours. After filtration to remove the solid, thefiltrate was concentrated to afford 83-F. LCMS-ESI⁺ (m/z): [M+H]⁺ found:263.

Step 6

A 100-mL round bottom flask was charged with 83-F (0.18 g, 0.69 mmol)and 83-G (0.324 g, 0.69 mmol) in ethanol (7 mL). Sodium bicarbonate(0.12 g, 1.4 mmol) in water (7 mL) was added to the reaction mixture.Then the reaction mixture was stirred at room temperature overnight. Themixture was diluted with EtOAc (50 mL) and washed with water. Theaqueous fractions were extracted with EtOAc, and the organic fractionswere combined, dried (Na₂SO₄), and concentrated. The crude 83-H was usedfor next step without further purification. LCMS-ESI⁺ (m/z): [M+H]⁺found: 591.

Step 7

A 100-mL round bottom flask was charged with 83-H (0.4 g, 0.68 mmol) in4 N HCl/dioxane (3.8 mL). Then the reaction mixture was stirred at roomtemperature for 1 hour. After concentration, 0.35 g intermediate wasobtained. The intermediate and DBU (0.51 g, 3.3 mmol) were dissolved intoluene (10 mL). The reaction mixture was heated to 110° C. withstirring for 1 hour. After concentration, the residue was purified byflash chromatography using hexanes-EtOAc as eluents to afford 83-I.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 431.

Step 8

A 100-mL round bottom flask was charged with 83-I (0.2 g, 0.47 mmol) inTHF (5 mL) and MeOH (5 mL). 1 N KOH (1.4 mL) was added to the reactionmixture. Then the reaction mixture was stirred at room temperature for 1hour. The reaction mixture was acidified by adding 1 N HCl (1.4 mL).After concentration, the residue was co-evaporated with toluene (3×).The crude acid, 2,4,6-trifluobenzylamine (0.14 g, 0.91 mmol),N,N-diisopropylethylamine (DIPEA) (0.29 g, 2.2 mmol) and HATU (0.35 g,0.91 mmol) were dissolved in DCM (10 mL). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted withEtOAc (100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl(2×) and dried over Na₂SO₄. After concentration, the crude was purifiedby column chromatography on silica gel with hexane-EtOAc to afford 83-J.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 560.

Step 9

A 50-mL rbf was charged with 83-J (0.18 g, 0.32 mmol) in TFA (3 mL). Thereaction mixture was stirred at room temperature for 30 minutes. Afterconcentration, the crude was purified by column chromatography on silicagel with EtOAc-MeOH to afford compound 83 as a white solid. ¹H-NMR (400MHz, Chloroform-d) δ 10.29 (d, J=6.1 Hz, 1H), 8.34 (s, 1H), 6.65 (dd,J=8.7, 7.5 Hz, 2H), 4.83 (s, 1H), 4.72-4.58 (m, 2H), 4.36-4.10 (m, 2H),4.05 (t, J=11.5 Hz, 1H), 2.97 (d, J=4.4 Hz, 1H), 2.49-2.08 (m, 3H),2.12-1.94 (m, 2H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −92.32 (ddd,J=225.6, 22.5, 9.1 Hz), −107.64˜−109.54 (m), −112.05 (t, J=7.0 Hz),−114.67 (d, J=226.7 Hz). LCMS-ESI⁺ (m/z): found: 470.

Example 84 Preparation of Compound 84(1S,2R,4S,12aR)-7-hydroxy-2-methyl-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL round bottom flask was charged with 84-A (1.6 g, 5.9 mmol) inDCM (20 mL). The reaction mixture was cooled to 0° C. Dess-Martinperiodinane (4.9 g, 11.7 mmol) was slowly added in. Then the reactionmixture was stirred at room temperature for 3 hours. Afterconcentration, the residue was purified by flash chromatography usinghexanes-EtOAc as eluents to afford 84-B. LCMS-ESI⁺ (m/z): [M+H]⁺ found:270.

Step 2

A 100-mL round bottom flask was charged with 84-B (1.3 g, 4.8 mmol) inTHF (30 mL). The reaction mixture was cooled to 0° C. Tebbe reagent (0.5M in toluene, 19.4 mL, 9.7 mmol) was slowly added in. Then the reactionmixture was stirred at room temperature for 2 hours. The mixture wascooled back to 0° C. Saturated NaHCO₃ (5 mL) was added drop wise toquench the reaction. The reaction mixture was stirred at roomtemperature for another 15 minutes and filtered through celite. Thefiltered cake was washed with DCM (2×). The combined filtrates wereconcentrated in vacuum and the residue was purified by flashchromatography using hexanes-EtOAc as eluents to afford 84-C. LCMS-ESI⁺(m/z): [M+H]⁺ found: 268.

Step 3

To a solution (purged with N₂) of 84-C (0.9 g, 3.4 mmol) in EtOH (20 mL)was added Pd/C (0.18 g). The mixture was stirred under H₂ for 3 hours.The mixture was filtered through celite and the filtrate wasconcentrated to afford 84-D. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 270.

Step 4

A 100-mL round bottom flask was charged with 84-D (0.9 g, 3.3 mmol) inTHF (6 mL). The reaction mixture was stirred at −78° C. 2.0 M LiBH₄ inTHF (13.2 mL, 26.4 mmol) was slowly added in. Then the reaction mixturewas warmed up and stirred at room temperature for 3 hours. Then thereaction mixture was diluted with EtOAc (100 mL) and added water slowly(H₂ evolution). After the two phases were separated, the aqueousfraction was extracted with EtOAc and the two organic fractions werecombined, washed with water, and dried over Na₂SO₄. After concentration,the residue was purified by flash chromatography using hexanes-EtOAc aseluents to afford 84-E. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 242.

Step 5

A 100-mL round bottom flask was charged with 84-E (0.4 g, 1.66 mmol),triphenylphosphine (0.96 g, 3.6 mmol) and phthalimide (0.39 g, 2.7 mmol)in THF (15 mL). Then the reaction mixture was cooled to 0° C. withstirring. DIAD (0.7 mL, 3.6 mmol) was slowly added to the reactionmixture. The reaction mixture was stirred at room temperature forovernight. After concentration, the residue was purified by flashchromatography using hexanes-EtOAc as eluents to afford 84-F. LCMS-ESI⁺(m/z): [M+H]⁺ found: 371.

Step 6

To a solution of 84-F (0.55 g, 1.5 mmol) EtOH (20 mL) was addedhydrazine monohydrate (0.3 mL). The reaction mixture was heated to 70°C. with stirring for 3 hours. After filtration to remove the solid, thefiltrate was concentrated to afford 84-G. LCMS-ESI⁺ (m/z): [M+H]⁺ found:241.

Step 7

A 100-mL round bottom flask was charged with 84-G (0.35 g, 1.4 mmol) and84-H (0.5 g, 1.4 mmol) in ethanol (10 mL). Sodium bicarbonate (0.24 g,2.8 mmol) in water (10 mL) was added to the reaction mixture. Then thereaction mixture was stirred at room temperature for overnight. Themixture was diluted with EtOAc (50 mL) and washed with water (2×). Theaqueous fractions were extracted with EtOAc, and the organic fractionswere combined, dried (Na₂SO₄), and concentrated. The crude 84-I was usedfor next step without further purification. LCMS-ESI⁺ (m/z): [M+H]⁺found: 583.

Step 8

A 100-mL rbf was charged with 84-I (0.84 g, 1.4 mmol) in 4 N HCl/dioxane (8.2 mL). Then the reaction mixture was stirred at roomtemperature for 1 hour. After concentration, 0.74 g intermediate wasobtained. The intermediate and DBU (1.1 g, 7.2 mmol) were dissolved intoluene (10 mL). The reaction mixture was heated to 110° C. withstirring for 1 hour. After concentration, the residue was purified byflash chromatography using hexanes-EtOAc as eluents to afford 84-J.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 409.

Step 9

A 100-mL round bottom flask was charged with 84-J (0.4 g, 0.98 mmol) inTHF (5 mL) and MeOH (5 mL). 1 N KOH (3.0 mL) was added to the reactionmixture. Then the reaction mixture was stirred at room temperature for 1hour. The reaction mixture was acidified by adding 1 N HCl (3.0 mL).After concentration, the residue was co-evaporated with toluene (3×).The crude acid, 2,4,6-trifluobenzylamine (0.32 g, 1.96 mmol),N,N-diisopropylethylamine (DIPEA) (0.63 g, 4.9 mmol) and HATU (0.74 g,1.9 mmol) were dissolved in DCM (10 mL). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted withEtOAc (100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl(2×) and dried over Na₂SO₄. After concentration, the crude was purifiedby column chromatography on silica gel with hexane-EtOAc to afford 84-K.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 538.

Step 10

A 50-mL round bottom flask was charged with 84-K (0.5 g, 0.93 mmol) inTFA (6 mL). The reaction mixture was stirred at room temperature for 30minutes. After concentration, the crude was purified by columnchromatography on silica gel with EtOAc-MeOH to afford compound 84.¹H-NMR (400 MHz, Chloroform-d) δ 10.37 (s, 1H), 8.28 (s, 1H), 6.65 (t,J=8.1 Hz, 2H), 4.80 (s, 1H), 4.77-4.52 (m, 3H), 4.08 (d, J=13.1 Hz, 1H),3.88 (d, J=12.3 Hz, 1H), 2.47 (d, J=3.2 Hz, 1H), 2.35 (s, 1H), 2.16(ddd, J=14.3, 11.2, 3.6 Hz, 1H), 1.93-1.57 (m, 3H), 1.29-1.19 (m, 1H),1.17 (d, J=7.0 Hz, 3H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −109.24,−111.98. LCMS-ESI⁺ (m/z): found: 448.

Example 85 Preparation of Compound 85(6aS,7R,11S)-1-hydroxy-2,13-dioxo-N-(2,4,6-trifluorobenzyl)-6,6a,7,8,9,10,11,13-octahydro-2H-7,11-methanopyrido[1′,2′:4,5]pyrazino[1,2-a]azepine-3-carboxamide

Step 1

A solution of 85-A (1100 mg, 3.855 mmol) in DMSO (6 mL) and water (0.75mL) was stirred at room temperature as N-iodosuccinmide (885 mg, 3.934mmol) was added. After 2 h, additional N-iodosuccinmide (88 mg, 0.391mmol) was added and the resulting mixture was stirred at roomtemperature for 1.5 h. The dark brown reaction mixture was diluted withEtOAc, and washed with a mixture of 10% aq. Na₂S₂O₃ solution and aq.NaHCO₃ solution (˜1:4 mixture) and then with water (with some brine).After the aqueous fractions were extracted with EtOAc, the organicfractions were combined, dried (Na₂SO₄), and concentrated. The residuewas purified by flash chromatography using hexanes-EtOAc as eluents toobtain 85-B. ¹H-NMR (400 MHz, CDCl₃) δ 7.51-7.44 (m, 2H), 7.33-7.17 (m,3H), 4.22-4.05 (m, 2H), 4.02-3.86 (m, 2H), 3.77 (d, J=5.3 Hz, 1H),3.54-3.44 (m, 1H), 3.27 (t, J=4.5 Hz, 1H), 2.75-2.66 (m, 1H), 2.30(dddd, J=14.8, 13.1, 7.2, 5.8 Hz, 1H), 2.14 (dddd, J=14.8, 13.0, 6.1,2.1 Hz, 1H), 1.97 (d, J=8.9 Hz, 1H), 1.58-1.46 (m, 1H), 1.45-1.34 (m,4H), 1.24 (t, J=7.1 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₈H₂₅1NO₃: 430.1; found: 430.0.

Step 2

A solution of 85-B (993 mg, 2.313 mmol), AIBN (305 mg, 1.857 mmol), andtributyltin hydride (1392 mg, 4.799 mmol) in toluene (15 mL) was stirredat 100° C. After 2 h, the reaction mixture was cooled to roomtemperature, diluted with EtOAc, and washed with water and brine. Afterthe aqueous fractions were extracted with EtOAc, the organic fractionswere combined, dried (Na₂SO₄), and concentrated. The residue waspurified by flash chromatography using hexanes-EtOAc as eluents toobtain 85-C. ¹H-NMR (400 MHz, CDCl₃) δ 7.57-7.49 (m, 2H), 7.32-7.23 (m,2H), 7.23-7.15 (m, 1H), 4.24-4.02 (m, 2H), 3.97 (q, J=6.7 Hz, 1H), 3.83(d, J=5.1 Hz, 1H), 3.48 (t, J=4.6 Hz, 1H), 3.19-3.04 (m, 1H), 2.58 (p,J=4.0 Hz, 1H), 2.30 (dddd, J=14.7, 13.1, 7.0, 4.5 Hz, 1H), 1.98 (d,J=11.2 Hz, 1H), 1.64 (tdd, J=13.3, 6.2, 2.6 Hz, 1H), 1.49-1.33 (m, 3H),1.37 (d, J=6.7 Hz, 3H), 1.32-1.26 (m, 1H), 1.23 (t, J=7.2 Hz, 3H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₈H₂₆NO₃: 304.2; found: 304.1.

Step 3

A mixture of 85-C (725 mg, 2.39 mmol) and 20% Pd(OH)₂/C (351 mg) in EtOH(25 mL) and 4 N HCl in dioxane (0.9 mL) was stirred under H₂ atmosphere.After 2 h, the reaction mixture was filtered, and the filtrate wasconcentrated. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₀H₁₈NO₃: 200.13;found: 200.1. After the residue was co-evaporated with toluene (×2), theresidue and Boc₂O (720 mg, 3.299 mmol) in THF (15 mL) was stirred atroom temperature as N,N-diisopropylethylamine (DIPEA) (1.2 mL, 6.889mmol) was added. After 1 h, the reaction mixture was diluted with waterand extracted with EtOAc (×2). After the organic extracts were washedwith water, the combined extracts were dried (Na₂SO₄) and concentrated.The residue was purified by flash using hexanes-EtOAc as eluents toobtain 85-D which appears to be a mixture of rotamers. ¹H-NMR (400 MHz,CDCl₃) δ 4.42-3.97 (m, 5H), 2.62 (d, J=5.6 Hz, 1H), 2.45-2.26 (m, 1H),2.25-2.15 (m, 1H), 1.80 (td, J=13.7, 6.7 Hz, 1H), 1.66 (dd, J=12.3, 6.6Hz, 2H), 1.55-1.70 (m, 2H), 1.47 (s, 2H), 1.42 (s, 7H), 1.28 (dt, J=9.5,7.1 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₅H₂₆NO₅: 300.2;found: 299.7.

Step 4

To a solution of 85-D (568 mg, 1.897 mmol) and pyridine (0.25 mL, 3.091mmol) in THF (5 mL) was added phenyl chlorothionoformate (0.3 mL, 2.169mmol) at 0° C., which produced insoluble material quickly. After −30 minat 0° C., additional pyridine (0.3 mL, 3.709 mmol) and phenylchlorothionoformate (0.3 mL, 2.169 mmol) were added. After 1.5 h at 0°C. and 1 h at room temperature, the mixture was concentrated, and theresidue was dissolved in EtOAc and water. After separation of twolayers, the organic fraction was washed with ˜0.1 N HCl, saturatedaqueous NaHCO₃, and brine. After the aqueous fractions were extractedwith EtOAc, the combined organic fractions were dried (Na₂SO₄), andconcentrated. The residue was purified by flash chromatography usingEtOAc/hexanes as eluents to afford 85-E. ¹H-NMR (400 MHz, CDCl₃) δ7.47-7.37 (m, 2H), 7.30 (t, J=6.9 Hz, 1H), 7.11 (dd, J=8.0, 4.0 Hz, 2H),5.54 (dt, J=9.0, 4.9 Hz, 1H), 4.50 (dt, J=9.8, 5.3 Hz, 1H), 4.35 (dd,J=21.4, 5.0 Hz, 1H), 4.30-4.14 (m, 2H), 2.71 (s, 1H), 2.54 (s, 1H),2.14-2.00 (m, 1H), 1.82 (m, 3H), 1.54 (m, 1H), 1.48 (s, 4.5H), 1.45 (s,4.5H), 1.30 (dt, J=9.4, 7.1 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₂H₃₀NO₆S: 436.2; found: 435.8.

Step 5

A mixture of 85-E (602 mg, 1.382 mmol), AIBN (182 mg, 1.108 mmol), andtributyltin hydride (608 mg, 2.096 mmol) in toluene (8 mL) was stirredat 100° C. After 1 h, the reaction mixture was concentrated and theresidue was dissolved in EtOAc before washing with water and brine.After the aqueous fractions were extracted with EtOAc, the combinedorganic fractions were dried (Na₂SO₄) and concentrated. The residue waspurified with flash chromatography using EtOAc/hexanes as eluents togive 85-F which appears to be a mixture of rotamers. ¹H-NMR (400 MHz,CDCl₃) δ 4.37-4.06 (m, 4H), 2.69-2.53 (m, 1H), 2.11 (m, 1H), 1.97 (m,0.65H), 1.93-1.80 (m, 1.35H), 1.54 (s, 5H), 1.46 (s, 3.15H), 1.42 (s,5.85H), 1.27 (m, 3H). LCMS-ESI⁺ (m/z): [M-C₄H₈+H]⁺ calculated forC₁₁H₁₈NO₄: 228.1; found: 227.9.

Step 6

85-F (420 mg) was repurified and the purified 85-F in THF (3 mL) wasstirred at 0° C. as 2.0 M LiBH₄ in THF (1.5 mL) was added. After 5 min,the mixture was stirred at room temperature for 17 h and additional 2.0M LiBH₄ in THF (1.5 mL) was added at room temperature. After 23 h atroom temperature, additional 2.0 M LiBH₄ in THF (3 mL) was added and theresulting mixture was stirred for ˜72 h. After the reaction mixture wasstirred at 0° C. as water was slowly added and further diluted withwater, the product was extracted with EtOAc (×2). The extracts werewashed with water, combined, dried (Na₂SO₄), and concentrated. Theresidue was purified by flash chromatography using hexane-EtOAc aseluents to give 85-G. ¹H-NMR (400 MHz, CDCl₃) δ 4.12 (t, J=5.3 Hz, 1H),3.99 (dd, J=12.0, 7.9 Hz, 1H), 3.85 (dd, J=8.0, 4.7 Hz, 1H), 3.73 (dd,J=11.9, 1.4 Hz, 1H), 2.28 (d, J=4.6 Hz, 1H), 1.90-1.73 (m, 2H),1.68-1.45 (m, 6H), 1.47 (s, 9H), 1.43-1.33 (m, 1H). LCMS-ESI⁺ (m/z):[M-C₄H₈+H]⁺ calculated for C₉H₁₆NO₃: 186.1; found: 186.0.

Step 7

A solution of 85-G (198 mg, 0.820 mmol), phthalimide (200 mg, 1.359mmol), and PPh₃ (488 mg, 1.861 mmol) in THF (10 mL) was stirred at 0° C.bath as DIAD (0.36 mL, 1.828 mmol) was added. After 30 min at 0° C., themixture was stirred at room temperature for 17 h. The reaction mixturewas concentrated and the residue was purified by flash chromatographyusing hexane-EtOAc as eluents to 85-H which appears to be a mixture ofrotamers. ¹H-NMR (400 MHz, CDCl₃) δ 7.82 (dd, J=5.4, 3.1 Hz, 2H), 7.69(dd, J=5.4, 3.1 Hz, 2H), 4.46 (s, 1H), 4.19 (m, 2H), 3.95 (s, 1H),2.31-2.14 (m, 1H), 2.05 (d, J=16.5 Hz, 1H), 1.84 (m, 2H), 1.79-1.70 (m,1H), 1.66 (m, 1H), 1.61-1.30 (m, 12H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₁H₂₇N₂O₄: 371.2; found: 370.8.

Step 8

To a solution of 85-H (270 mg, 0.729 mmol) in EtOH (12 mL) was addedhydrazine hydrate (0.145 mL, 3.083 mmol) at room temperature and theresulting solution was stirred at 70° C. After 1.5 h, the mixture wascooled to 0° C. and diluted with ether (30 mL) before stirring for 1 hat 0° C. The mixture was filtered and the filtrate was concentrated. Theresidue was dissolved in CH₂Cl₂ and filtered to remove some insolublematerial. The resulting filtrate was concentrated. The residue, combinedwith 85-I (257 mg, 0.742 mmol), and NaHCO₃ (131 mg, 1.559 mmol) in water(3 mL) and EtOH (3 mL) was stirred at room temperature. After 1 h, themixture was diluted with water and extracted with EtOAc (×2). After theextracts were washed with water, the organic extracts were combined,dried (Na₂SO₄), and concentrated. To a solution of the residue in CH₂Cl₂(2 mL) was added 4 N HCl in dioxane (6 mL). After 1.5 h at roomtemperature, the solution was concentrated and co-evaporated withtoluene. A mixture of the residue and DBU (0.6 mL, 4.012 mmol) intoluene (5 mL) was stirred at 100° C. bath. After 1 h, additional DBU(0.3 mL, 2.006 mmol) was added and the mixture was stirred another 1 hat 100° C. After the mixture was concentrated, the residue was purifiedby flash chromatography using EtOAc-20% MeOH/EtOAc as eluents to give85-J. ¹H-NMR (400 MHz, CDCl₃) δ 8.08 (s, 1H), 7.71-7.62 (m, 2H),7.36-7.29 (m, 2H), 7.29-7.23 (m, 1H), 5.44 (d, J=9.8 Hz, 1H), 5.10 (d,J=9.8 Hz, 1H), 4.44-4.28 (m, 3H), 4.23 (t, J=13.0 Hz, 1H), 3.99 (ddt,J=10.2, 6.3, 3.6 Hz, 2H), 2.44-2.36 (m, 1H), 2.29 (dt, J=11.6, 5.3 Hz,1H), 1.84 (dt, J=10.8, 5.3 Hz, 2H), 1.77-1.61 (m, 3H), 1.57 (d, J=11.7Hz, 1H), 1.48 (ddd, J=20.9, 12.3, 5.5 Hz, 1H), 1.38 (t, J=7.1 Hz, 3H).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₄H₂₇N₂O₅: 423.2; found: 423.3.

Step 9

A mixture of 85-J (214 mg, 0.507 mmol) in THF (4 mL) and MeOH (4 mL) wasstirred at room temperature as 1 N KOH (1.1 mL) was added. After 30 min,the reaction mixture was concentrated to −1 mL, acidified with 1 N HCl(˜1.2 mL), and diluted with brine before extraction with CH₂Cl₂ (20mL×2). The combined extracts were dried (Na₂SO₄) and concentrated toobtain the crude acid. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₃N₂O₅: 395.2; found: 395.3.

A mixture of the crude acid (199 mg, 0.505 mmol), 2,4,6-trifluorobenzylamine (130 mg, 0.807 mmol), and HATU (304 mg, 0.800 mmol) in CH₂Cl₂ (6mL) was stirred at room temperature as N,N-diisopropylethylamine (DIPEA)(0.62 mL, 3.559 mmol) was added. After 30 min, the reaction mixture wasconcentrated and the residue was dissolved in EtOAc, washed withsaturated aqueous NH₄Cl (×2), saturated aqueous NaHCO₃ (×2), and brine.After the aqueous fractions were extracted with EtOAc, two organicfractions were combined, dried (Na₂SO₄) and concentrated. The residuewas purified by flash using EtOAc-20% MeOH/EA as eluents to obtain 85-K.¹H-NMR (400 MHz, CDCl₃) δ 10.40 (t, J=5.7 Hz, 1H), 8.42 (s, 1H),7.68-7.54 (m, 2H), 7.33 (ddd, J=7.7, 6.3, 1.5 Hz, 2H), 7.30-7.26 (m,1H), 6.74-6.60 (m, 2H), 5.37 (d, J=10.0 Hz, 1H), 5.17 (d, J=10.0 Hz,1H), 4.76-4.57 (m, 2H), 4.46 (dd, J=6.0, 4.3 Hz, 1H), 4.34 (t, J=12.4Hz, 1H), 4.07 (dd, J=12.4, 3.6 Hz, 1H), 3.91 (dt, J=12.4, 3.9 Hz, 1H),2.52-2.44 (m, 1H), 2.32 (dd, J=11.8, 6.2 Hz, 1H), 1.92 (dt, J=10.7, 5.4Hz, 1H), 1.83-1.70 (m, 3H), 1.67 (d, J=11.7 Hz, 1H), 1.52 (dddt, J=25.5,17.0, 11.8, 5.3 Hz, 2H). ¹⁹F-NMR (376 MHz, CDCl₃) δ −109.15 (dq, J=15.0,7.5, 7.1 Hz, 1F), −111.85 (t, J=6.8 Hz, 2F). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₉H₂₇F₃N₃O₄: 538.2; found: 538.3.

Step 10

85-K (187 mg, 0.348 mmol) was dissolved in trifluoroacetic acid (3 mL)at room temperature and stirred at room temperature. After 1 h, thesolution was concentrated and the residue was dissolved in CH₂Cl₂. Afterthe solution was washed with 0.1 N HCl, the aqueous fraction wasextracted with CH₂Cl₂ (×2). The organic fractions were combined, dried(Na₂SO₄), and concentrated. The residue was purified by flashchromatography using CH₂Cl₂-20% MeOH in CH₂Cl₂ as eluents to obtain 150mg (96%) of compound 85. Compound 85 was further purified byrecrystallization from methanol (10 mL) to give compound 85. ¹H-NMR (400MHz, CDCl₃) δ 12.09 (s, 1H), 10.39 (t, J=5.7 Hz, 1H), 8.36 (s, 1H),6.74-6.48 (m, 2H), 4.64 (d, J=5.7 Hz, 2H), 4.59 (dd, J=6.1, 4.4 Hz, 1H),4.36-4.18 (m, 2H), 4.12 (dt, J=12.4, 4.1 Hz, 1H), 2.68-2.47 (m, 1H),2.25-2.10 (m, 1H), 2.10-1.98 (m, 1H), 1.98-1.66 (m, 4H), 1.66-1.48 (m,2H). ¹⁹F-NMR (376 MHz, CDCl₃) δ −109.23 (ddd, J=15.1, 8.6, 6.0 Hz, 1F),−112.02 (t, J=6.9 Hz, 2F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₁F₃N₃O₄: 448.2; found: 448.3.

Example 86 Preparation of Compound 86(1R,3S,4R,12aS)-7-hydroxy-3-methyl-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A solution of 86-A (10.160 g, 39.48 mmol) in DMSO (52 mL) and water (6.5mL) was stirred at room temperature as N-iodosuccinmide (8.888 g, 39.50mmol) was added. After 30 min, the dark brown reaction mixture wasdiluted with EtOAc, and washed with saturated aqueous NaHCO₃ solution,10% aqueous Na₂S₂O₃ solution], and brine. After the aqueous fractionswere extracted with EtOAc, the organic fractions were combined, dried(Na₂SO₄), and concentrated. The residue was purified by flashchromatography using hexanes-EtOAc as eluents to obtain 86-B as a whitesolid. ¹H-NMR (400 MHz, CDCl₃) δ 7.33-7.19 (m, 5H), 4.25-4.12 (m, 1H),3.79 (q, J=1.6 Hz, 1H), 3.72 (q, J=6.5 Hz, 1H), 3.51 (s, 1H), 3.47 (s,3H), 3.31 (dd, J=3.9, 1.6 Hz, 1H), 2.76-2.69 (m, 1H), 2.13 (ddd, J=14.3,7.8, 1.7 Hz, 1H), 2.08-1.97 (m, 1H), 1.91 (dtd, J=14.1, 4.0, 1.5 Hz,1H), 1.42 (d, J=6.5 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₆H₂₁INO₃: 402.1; found: 402.0.

Step 2

A solution of 86-B (12.468 g, 31.07 mmol), azobisisobutyronitrile (AIBN)(4.082 g, 24.86 mmol), and tributyltin hydride (18.047 g 62.22 mmol) intoluene (150 mL) was stirred at 100° C. After 30 min, the reactionmixture was cooled to room temperature, diluted with EtOAc, and washedwith water and brine. After the aqueous fractions were extracted withEtOAc, the organic fractions were combined, dried (Na₂SO₄), andconcentrated. The residue was purified by flash chromatography twiceusing hexanes-EtOAc as eluents to obtain 86-C. ¹H-NMR (400 MHz, CDCl₃) δ7.39-7.31 (m, 2H), 7.31-7.24 (m, 2H), 7.24-7.17 (m, 1H), 4.11 (s, 1H),3.72 (s, 1H), 3.49 (s, 3H), 3.33 (d, J=3.4 Hz, 1H), 3.27 (d, J=6.4 Hz,1H), 2.65-2.51 (m, 1H), 1.92 (ddd, J=13.6, 6.8, 2.4 Hz, 1H), 1.69-1.50(m, 2H), 1.47 (d, J=10.1 Hz, 1H), 1.41 (d, J=6.6 Hz, 3H), 1.21-1.07 (m,1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₆H₂₂NO₃: 276.2; found:276.1.

Step 3

A mixture of 86-C (4.187 g, 15.21 mmol) and 20/o Pd(OH)₂/C (1.022 g) inEtOH (100 mL) and 4 N HCl in dioxane (5.7 mL) was stirred under H₂atmosphere. After 1.5 h, the reaction mixture was filtered, and thefiltrate was concentrated. After the residue was co evaporated withtoluene, the residue was used for the next step. LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₈H₁₄NO₃: 172.1; found: 172.1.

After the residue was co-evaporated with toluene, the residue and Boc₂O(5.712 g, 26.17 mmol) in THF (45 mL) was stirred at room temperature asN,N-diisopropylethylamine (DIPEA) (8 mL, 45.93 mmol) was added. After 30min, the reaction mixture was diluted with water and extracted withEtOAc (×2). After the organic extracts were washed with water, thecombined extracts were dried (Na₂SO₄) and concentrated. The residue waspurified by flash chromatography using hexanes-EtOAc as eluents toobtain 86-D. ¹H NMR spectrum suggests a mixture of rotamers. ¹H-NMR (400MHz, CDCl₃) δ 4.20 (d, J=7.6 Hz, 1H), 4.19-4.10 (m, 2H), 4.08 (d, J=3.5Hz, 1H), 3.72 (s, 3H), 2.74 (d, J=5.6 Hz, 1H), 1.97 (ddd, J=13.6, 6.9,2.8 Hz, 1H), 1.88-1.78 (m, 1H), 1.79-1.50 (m, 1H), 1.46 (s, 3H), 1.38(s, 6H), 1.31 (d, J=13.3 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₃H₂₂NO₅: 272.2; found: 271.6.

Step 4

A solution of 86-D (1659 mg, 6.115 mmol) in CH₂Cl₂ (35 mL) was stirredat 0° C. bath as Dess-Martin periodinane (5.183 g, 12.22 mmol) was addedin portions. After 5 min, the mixture was stirred at room temperature.After 2 h, the reaction mixture was cooled in an ice bath, quenched withwater, and filtered. The filtrate was washed with saturated NaHCO₃,dried (Na₂SO₄), and concentrated. The residue was purified by flashchromatography using hexanes-EtOAc as eluents to give 86-E. ¹H NMRspectrum suggests two rotamers. ¹H-NMR (400 MHz, CDCl₃) δ 4.43 (d, J=3.8Hz, 0.5H), 4.39 (s, 1H), 4.26 (s, 0.5H), 3.75 (s, 3H), 3.10 (s, 1H),2.24 (d, J=4.5 Hz, 0.5H), 2.19 (d, J=4.4 Hz, 0.5H), 2.12 (d, J=4.4 Hz,0.5H), 2.07 (d, J=4.2 Hz, 0.5H), 2.01 (dd, J=4.5, 2.2 Hz, 0.5H), 1.98(dt, J=4.3, 1.9 Hz, 0.5H), 1.80 (s, 0.5H), 1.77 (s, 0.5H), 1.46 (s,4.5H), 1.40 (d, J=2.8 Hz, 4.5H). LCMS-ESI⁺ (m/z): [M-C₄H₈+H]⁺ calculatedfor C₉H₁₂NO₅: 214.1; found: 213.8.

Step 5

A solution of 86-E (528 mg, 1.961 mmol) in THF (12 mL) was stirred at 0°C. as 0.5 M solution of Tebbe reagent in toluene (7.9 mL, 3.95 mmol) wasadded dropwise. After addition, the brown solution was allowed to warmto room temperature slowly and was stirred at room temperature for 2.5h. The reaction mixture was stirred at 0° C. bath as the reaction wasquenched carefully by the addition of saturated aqueous NaHCO₃ solution.After the mixture was diluted with CH₂Cl₂ and stirred at roomtemperature for 15 minutes, the resulting mixture was filtered throughcelite pad and the filter cake was washed with CH₂Cl₂. After the twofractions in the filtrate were separated, the aq. fraction was extractedwith CH₂Cl₂, and the organic fractions were combined, dried (Na₂SO₄),and concentrated. The residue was purified by flash chromatography usinghexanes-EtOAc as eluents to give 86-F. ¹H NMR spectrum suggests tworotamers. ¹H-NMR (400 MHz, CDCl₃) δ 5.13 (s, 0.6H), 5.04 (s, 0.4H),4.82-4.71 (m, 1H), 4.55 (s, 0.6H), 4.43 (s, 0.4H), 4.29 (d, J=3.7 Hz,0.4H), 4.24 (d, J=3.7 Hz, 0.6H), 3.71 (s, 3H), 2.84 (s, 1H), 2.14 (m,2H), 1.75 (s, 0.6H), 1.74-1.70 (s, 0.4H), 1.55 (m, 1H), 1.45 (s, 3.6H),1.37 (s, 5.4H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₄H₂₂NO₄: 268.2;found: 267.6.

Step 6

A mixture of 86-F (333 mg, 1.246 mmol) and 20/o Pd(OH)₂/C (53 mg) inEtOH (5 mL) was stirred under H₂ atmosphere. After 30 min, the mixturewas filtered and the filtrate was concentrated to give 86-G. ¹H NMRspectrum suggests two rotamers. ¹H-NMR (400 MHz, CDCl₃) δ 4.20 (m, 1H),4.08 (m, 1H), 3.71 (two s, 3H), 2.68 (m, 1H), 2.06 (m, 1H), 1.80-1.63(m, 2H), 1.63-1.51 (m, 1H), 1.44 (s, 4H), 1.38 (s, 5H), 1.13 (m, 3H),0.92 (m, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₄H₂₄NO₄: 270.2;found: 269.7.

Step 7

A solution of 86-G (336 mg, 1.482 mmol) in THF (5 mL) was stirred at 0°C. as 2.0 M LiBH₄ in THF (1.5 mL) was added. After 5 min, the mixturewas stirred at room temperature. After 2 h, additional 2.0 M LiBH₄ inTHF (1.5 mL) was added. After 21 h at room temperature, additional 2.0 MLiBH₄ in THF (3 mL) was added. After 3 h at room temperature, thesolution was heated at 35° C. for 18 h. The reaction mixture was cooledto 0° C. and quenched carefully with water. After the mixture wasextracted with EtOAc (×2), the two organic fractions were washed withwater, combined, dried (Na₂SO₄), and concentrated. The residue waspurified by flash chromatography using hexanes-EtOAc to give 86-H.¹H-NMR (400 MHz, CDCl₃) δ 4.95-4.09 (br, 1H), 4.05 (s, 1H), 3.82 (dd,J=11.5, 7.7 Hz, 1H), 3.76-3.69 (m, 1H), 3.66 (d, J=11.5 Hz, 1H), 2.45(d, J=4.1 Hz, 1H), 2.03 (dqdd, J=11.4, 7.0, 4.5, 2.6 Hz, 1H), 1.77-1.57(m, 2H), 1.48 (dd, J=10.1, 1.8 Hz, 1H), 1.45 (s, 9H), 1.00 (d, J=6.9 Hz,3H), 0.93 (ddd, J=13.2, 4.7, 2.6 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₁₃H₂₄NO₃: 242.2; found: 241.7.

Step 8

A solution of 86-H (218 mg, 0.903 mmol), phthalimide (218 mg, 1.482mmol), and PPh₃ (535 mg, 2.040 mmol) in THF (10 mL) was stirred at 0° C.bath as DIAD (0.40 mL, 2.032 mmol) was added. After 10 min at 0° C., themixture was stirred at room temperature for 19 h. The reaction mixturewas concentrated and the residue was purified by flash chromatographyusing hexane-EtOAc as eluents to give 86-I. ¹H NMR suggests tworotamers. ¹H-NMR (400 MHz, CDCl₃) δ 7.82 (dt, J=7.3, 3.6 Hz, 2H), 7.70(d, J=5.3 Hz, 2H), 4.53-4.26 (m, 1H), 4.26-3.89 (m, 2H), 3.89-3.65 (m,1H), 2.28 (m, 1H), 2.04 (m, 1H), 1.82-1.65 (m, 2H), 1.66-1.43 (m, 7H),1.38 (s, 4H), 1.19-1.01 (m, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₂₇N₂O₄: 371.2; found: 370.8.

Step 9

To a solution of 86-I (319 mg, 0.861 mmol) in EtOH (12 mL) was addedhydrazine hydrate (0.17 mL, 3.494 mmol) at room temperature and theresulting solution was stirred at 70° C. bath. After 1.5 h, the mixturewas cooled to 0° C. and diluted with ether (25 mL) before stirring for 1h at 0° C. The mixture was filtered and the filtrate was concentrated.The residue was dissolved in CH₂Cl₂ and filtered to remove someinsoluble material. The resulting filtrate was concentrated to givecrude amine. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₃H₂₅N₂O₂: 241.2;found: 240.9.

After the crude amine was co-evaporated with toluene, a mixture of thecrude amine, 85-I (300 mg, 0.866 mmol), and NaHCO₃ (150 mg, 1.845 mmol)in water (3 mL) and EtOH (3 mL) was stirred at room temperature. After 2h, the mixture was diluted with water and extracted with EtOAc (×2).After the extracts were washed with water, the organic extracts werecombined, dried (Na₂SO₄), and concentrated. To a solution of the residuein CH₂Cl₂ (2 mL) was added 4 N HCl in dioxane (6 mL). After 1.5 h atroom temperature, the solution was concentrated and co-evaporated withtoluene. A mixture of the residue and DBU (0.65 mL, 4.347 mmol) intoluene (6 mL) was stirred at 100° C. After 1 h, additional DBU (0.65mL, 4.347 mmol) was added and the mixture was stirred at 100° C.Additional DBU (0.65 mL, 4.347 mmol) was added after 1 h and the mixturewas stirred another 2.5 h at 100° C. The mixture was diluted with CH₂Cl₂and washed with water containing 3 mL of 1 N HCl. The organic fractionwas dried (Na₂SO₄) and concentrated. The residue was purified by flashchromatography using EtOAc-20% MeOH/EtOAc as eluents to give 86-J.¹H-NMR (400 MHz, CDCl₃) δ 8.09 (s, 1H), 7.70-7.62 (m, 2H), 7.37-7.27 (m,3H), 5.48 (d, J=9.9 Hz, 1H), 5.16 (d, J=9.9 Hz, 1H), 4.53 (s, 1H), 4.38(m, 2H), 4.11 (m, 1H), 3.97 (dd, J=12.2, 3.0 Hz, 1H), 3.88 (dt, J=12.2,3.0 Hz, 1H), 2.63 (d, J=4.2 Hz, 1H), 2.28 (qd, J=7.2, 3.1 Hz, 1H),2.00-1.88 (m, 1H), 1.80-1.56 (m, 2H), 1.39 (t, J=7.1 Hz, 3H), 1.07 (d,J=6.9 Hz, 3H), 1.04 (dd, J=5.0, 2.5 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₄H₂₇N₂O₅: 423.2; found: 423.2.

Step 10

A mixture of 86-J (83 mg, 0.196 mmol) in THF (2 mL) and EtOH (2 mL) wasstirred at room temperature as 1 N KOH (0.4 mL) was added. After 30 min,the reaction mixture was diluted with water and washed with CH₂Cl₂.After the aqueous fraction was acidified with 1 N HCl 0.45 mL), theproduct was extracted with CH₂Cl₂ (×2). The combined extracts were dried(Na₂SO₄) and concentrated to obtain the crude acid. LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₂H₂₃N₂O₅: 395.2; found: 395.2.

A mixture of the crude acid (69 mg, 0.175 mmol), 2,4,6-trifluorobenzylamine (42 mg, 0.261 mmol), and HATU (106 mg, 0.279 mmol) in CH₂Cl₂ (3mL) was stirred at room temperature as N,N-diisopropylethylamine (DIPEA)(0.25 mL, 1.435 mmol) was added. After 30 min, the reaction mixture wasconcentrated and the residue was dissolved in EtOAc, washed withsaturated aqueous NH₄Cl (×2), saturated aqueous NaHCO₃ (×2), and brine.After the aqueous fractions were extracted with EtOAc, two organicfractions were combined, dried (Na₂SO₄) and concentrated. The residuewas purified by flash chromatography using EtOAc-20% MeOH/EtOAc aseluents to obtain 86-K. ¹H-NMR (400 MHz, CDCl₃) δ 10.40 (t, J=5.7 Hz,1H), 8.40 (s, 1H), 7.66-7.51 (m, 2H), 7.36-7.29 (m, 2H), 7.29-7.23 (m,1H), 6.71-6.61 (m, 2H), 5.36 (d, J=10.0 Hz, 1H), 5.18 (d, J=10.0 Hz,1H), 4.73-4.58 (m, 2H), 4.53 (s, 1H), 4.22-4.11 (m, 1H), 4.03 (dd,J=12.4, 3.1 Hz, 1H), 3.81 (dt, J=12.3, 3.1 Hz, 1H), 2.68-2.59 (m, 1H),2.29 (dddd, J=11.4, 7.1, 4.7, 2.4 Hz, 1H), 1.94 (ddd, J=13.5, 11.2, 4.6Hz, 1H), 1.88-1.67 (m, 2H), 1.06 (d, J=7.0 Hz, 3H), 1.03-1.09 (m, 1H).¹⁹F-NMR (376 MHz, CDCl₃) δ −109.14 (ddd, J=15.2, 8.7, 6.2 Hz, 1F),−111.86 (t, J=7.0 Hz, 2F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₉H₂₇F₃N₃O₄: 538.2; found: 538.1.

Step 11

86-K (61 mg, 0.113 mmol) was dissolved in trifluoroacetic acid (2 mL)and stirred at room temperature. After 1 h, the solution wasconcentrated and the residue was dissolved in CH₂Cl₂. After the solutionwas washed with 0.1 N HCl, the aqueous fraction was extracted withCH₂Cl₂ (×2). The organic fractions were combined, dried (Na₂SO₄), andconcentrated. The residue was purified by flash chromatography usingCH₂Cl₂-20% MeOH in CH₂Cl₂ as eluents to obtain compound 86. ¹H-NMR (400MHz, CDCl₃) δ 12.02 (s, 1H), 10.40 (t, J=5.7 Hz, 1H), 8.35 (s, 1H), 6.63(t, J=8.1 Hz, 2H), 4.62 (d, J=5.7 Hz, 2H), 4.59 (s, 1H), 4.22 (dd,J=12.2, 3.5 Hz, 1H), 4.13 (t, J=11.9 Hz, 1H), 4.05 (dt, J=12.0, 3.1 Hz,1H), 2.77-2.70 (m, 1H), 2.31 m, 1H), 2.09-1.93 (m, 1H), 1.93-1.81 (m,2H), 1.10 (ddd, J=13.9, 5.0, 2.1 Hz, 1H), 1.02 (d, J=6.9 Hz, 3H).¹⁹F-NMR (376 MHz, CDCl₃) δ −109.22 (ddd, J=15.1, 8.7, 6.1 Hz, 1F),−112.05 (t, J=6.9 Hz, 2F). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₁F₃N₃O₄: 448.2; found: 448.3.

Example 87 Preparation of cis-5-aminotetrahydro-2H-pyran-3-ol

Step 1

A solution of benzyl (5-oxotetrahydro-2H-pyran-3-yl)carbamate (740 mg,3.0 mmol) and cerium(III) chloride heptahydrate (1.12 g, 3.0 mmol) in 20mL methanol was cooled to 0° C. and sodium borohydride (120 mg, 3.2mmol) was then added portionwise. The reaction mixture was allowed tostir at 0° C. for 45 minutes and then quenched by slow addition of 1 mLacetone followed by 3 hours stirring at room temperature. The reactionmixture was partitioned between water and dichloromethane and theaqueous phase extracted into dichloromethane followed by 2-butanol. Thecombined organic phases were dried over magnesium sulfate, filtered,concentrated, and the residue purified by flash chromatography (0-100%EtOAc/hexanes) to afford the desired cis-benzyl((3R,5S)-5-hydroxytetrahydro-2H-pyran-3-yl)carbamate. ¹H-NMR (400 MHz,Chloroform-d) δ 7.39-7.26 (m, 5H), 6.06 (br s, 1H), 5.07 (s, 2H),3.86-3.70 (m, 2H), 3.69-3.47 (m, 4H), 2.00-1.89 (m, 1H), 1.76 (d, J=13.5Hz, 1H). The undesired trans-isomer was also isolated.

Step 2

To a solution of cis-benzyl((3R,5S)-5-hydroxytetrahydro-2H-pyran-3-yl)carbamate (290 mg, 1.16 mmol)in 5 mL 1:1 DCM:EtOH was added 10 wt % Pd/C (255 mg). This mixture wasstirred under balloon pressure hydrogen for 18 hours and palladiumremoved by filtration thru celite with ethanol rinse. Upon concentrationof filtrate, the cis-5-aminotetrahydro-2H-pyran-3-ol was afforded andcarried on as crude.

Example 88 Preparation of Compound 88′(2R,5S,13aR)—N-(3-chloro-2-fluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 88 was prepared in a similar manner to compound 15 using(3-chloro-2-fluorophenyl)methanamine in place of(4-fluorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ 10.43 (brs, 1H), 8.34 (br s, 1H), 7.32-7.24 (m, 2H), 7.02 (t, J=7.9 Hz, 1H), 5.36(d, J=9.4 Hz, 1H), 5.30 (s, 2H), 4.70 (d, J=6.0 Hz, 3H), 4.24 (d, J=12.0Hz, 1H), 4.00 (dd, J=12.7, 9.5 Hz, 1H), 2.18-1.96 (m, 4H), 1.96-1.83 (m,1H), 1.60 (dt, J=12.4, 3.1 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₁H₁₉ClFN₃O₅: 448.11; found: 448.2.

Example 89 Preparation of Compound 89(2R,5S,13aR)—N-(2,5-difluorobenzyl)-8-hydroxy-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide

Compound 89 was prepared in a similar manner to compound 15 using(2,5-difluorophenyl)methanamine in place of (4-fluorophenyl)methanamine.¹H-NMR (400 MHz, Chloroform-d) δ 10.32 (t, J=5.8 Hz, 1H), 8.31 (br s,1H), 7.15-6.89 (m, 2H), 6.86 (d, J=8.5 Hz, 1H), 5.40 (d, J=9.3 Hz, 1H),5.24 (s, 1H), 4.67-4.51 (m, 3H), 4.35-4.28 (m, 1H), 3.99-3.90 (m, 1H),2.16-1.85 (m, 5H), 1.60-1.50 (m, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₁H₁₉F₂N₃O₅: 432.14; found: 432.2.

Example 90 Preparation of Compound 90(1R,4S,12aR)—N-(3-chloro-2-fluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 90 was prepared in a similar manner to compound 41 using(3-chloro-2-fluorophenyl)methanamine in place of(2,4,6-trifluorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ9.22 (s, 1H), 8.79 (s, 1H), 7.39-7.28 (m, 2H), 7.06 (t, J=8.0 Hz, 1H),4.89 (s, 1H), 4.70-4.56 (m, 3H), 4.06-3.83 (m, 2H), 3.04-2.88 (m, 1H),2.77 (s, 1H), 1.97-1.58 (m, 6H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₁H₁₉ClFN₃O₄: 432.11; found: 432.2.

Example 91 Preparation of Compound 91(1R,4S,12aR)-7-hydroxy-6,8-dioxo-N-(2,3,4-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 91 was prepared in a similar manner to compound 41 using(2,3,4-trifluorophenyl)methanamine in place of(2,4,6-trifluorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ10.25 (s, 1H), 8.45 (s, 1H), 7.10 (d, J=5.1 Hz, 1a), 6.90 (d, J=8.7 Hz,1H), 4.89 (s, 1H), 4.63 (s, 2H), 4.22 (d, J=11.6 Hz, 1H), 3.93-3.73 (m,2H), 2.71 (s, 1H), 1.97-1.57 (m, 6H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₁H₁₈F₃N₃O₄: 434.13; found: 434.2.

Example 92 Preparation of Compound 92(1R,4S,12aR)—N-(4-chloro-2-fluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 92 was prepared in a similar manner to compound 41 using(4-chloro-2-fluorophenyl)methanamine in place of(2,4,6-trifluorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d) δ10.28 (s, 1H), 8.41 (s, 1H), 7.29 (s, 1H), 7.11-6.95 (m, 2H), 4.85 (s,1H), 4.57 (s, 2H), 4.22 (d, J=10.2 Hz, 1a), 3.81 (q, J=13.9, 13.1 Hz,2H), 2.68 (s, 1H), 1.99-1.50 (m, 6H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculatedfor C₂₁H₁₉ClFN₃O₄: 432.11; found: 432.2.

Example 93 Preparation of Compound 93 (R,4S,12aR)—N-(2-chloro-4,6-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 5 mL microwave vial was charged with2-bromo-1-chloro-3,5-difluorobenzene (540 mg, 2.4 mmol), cuprous cyanide(436 mg, 4.87 mmol), tetrakis(triphenylphosphine)palladium (63 mg, 0.05mmol), sealed, and evacuated/backfilled with nitrogen. To this was added5 mL degassed DMF. The sealed vessel was heated at 110° C. for 18 hours,diluted with ethyl acetate, and washed sequentially with twice 9:1NH₄OH:NH₄Cl_((aq)), twice 5% LiCl_((aq)), and brine. The organic phasewas then dried over magnesium sulfate, filtered, and concentrated. Thecrude residue was purified by flash chromatography (100% hexanes) toafford 2-chloro-4,6-difluorobenzonitrile. ¹H-NMR (400 MHz, Chloroform-d)δ 7.13 (dt, J=8.0, 1.9 Hz, 1H), 6.93 (td, J=8.5, 2.3 Hz, 1H).

Step 2

To a solution of 2-chloro-4,6-difluorobenzonitrile (210 mg, 1.2 mmol) in2.4 mL THF was added a 2M solution of borane-DMS in THF (0.6 mL). Thisreaction mixture was allowed to stir at refluxing temperature for 18hours resulting in a loss of all solvent. The residue was re-dissolvedin 3 mL THF, cooled to 0° C., a 6M solution of HCl_((aq)) was carefullyadded, and the mixture returned to reflux for 30 minutes. The reactionmixture was once again cooled to 0° C. and treated with 4M NaOH_((aq)).The aqueous phase was extracted with DCM, combined organic phases driedover magnesium sulfate, filtered, and concentrated. The crude residuewas purified by flash chromatography (0-10% MeOH/DCM) to afford(2-chloro-4,6-difluorophenyl)methanamine. ¹H-NMR (400 MHz, Chloroform-d)δ 6.95 (dt, J=8.3, 2.1 Hz, 1H), 6.76 (td, J=9.4, 2.5 Hz, 1H), 3.94 (d,J=1.9 Hz, 2H).

Steps 3 and 4

A solution of 93-A (74 mg, 0.11 mmol),(2-chloro-4,6-difluorophenyl)methanamine (48.5 mg, 0.27 mmol), HATU (100mg, 0.26 mmol), and N,N-diisopropylethylamine (0.1 mL, 0.57 mmol) in 1mL dichloromethane was stirred at room temperature for one hour at whichpoint complete disappearance of 93-A and formation of 93-B was observedby LCMS. TFA (0.65 M) was added and the mixture was stirred at roomtemperature for one hour, at which point 1 mL DMF was added. Thereaction mixture and then concentrated and purified by preparative HPLC(ACN/H₂O+0.1% TFA) to afford compound 93. ¹H-NMR (400 MHz, DMSO-d₆) δ10.41 (t, J=5.7 Hz, 1H), 8.33 (s, 1H), 7.41-7.26 (m, 2H), 4.72-4.57 (m,3H), 4.43 (dd, J=12.5, 3.6 Hz, 1H), 3.94 (t, J=12.4 Hz, 2H), 3.77 (dd,J=12.4, 3.6 Hz, 3H), 1.87-1.67 (m, 3H), 1.67-1.45 (m, 2H), 1.43 (d,J=10.4 Hz, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₈ClF₂N₃O₄:450.10; found: 450.2.

Example 94 Preparation of Compound 94(1R,4S,12aR)—N-benzyl-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Compound 94 was prepared in a similar manner to compound 41 usingphenylmethanamine in place of (2,4,6-trifluorophenyl)methanamine. ¹H-NMR(400 MHz, Chloroform-d) δ 10.37 (s, 1H), 8.26 (s, 1H), 7.37-7.19 (m,5H), 4.55 (d, J=4.8 Hz, 1H), 4.34 (d, J=5.7 Hz, 1H), 4.23 (d, J=9.8 Hz,1H), 4.09 (d, J=28.2 Hz, 1H), 3.78 (d, J=10.9 Hz, 1H), 3.64 (d, J=13.2Hz, 1H), 3.14-3.01 (m, 1H), 1.91-1.49 (m, 4H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₁H₂₁N₃O₄: 380.16; found: 380.2.

Example 95 Preparation of chiral tert-butyl3-((1,3-dioxoisoindolin-2-yl)methyl)-2-azabicyclo[2.1.1]hexane-2-carboxylates95-A and 95-B

Step 1

To a 0° C. solution of racemic tert-butyl3-(hydroxymethyl)-2-azabicyclo[2.1.1]hexane-2-carboxylate (285 mg, 1.34mmol), triphenylphosphine (425 mg, 1.62 mmol), and phthalimide (240 mg,1.62 mmol) in 9 mL THF was added dropwise a solution of diisopropylazodicarboxylate (0.35 mL, 1.8 mmol) in 1 ml THF. The reaction mixturewas warmed to room temperature, stirred for 90 minutes, concentratedonto silica, and purified by flash chromatography (0-25% EtOAc/hexanes)to afford tert-butyl3-((1,3-dioxoisoindolin-2-yl)methyl)-2-azabicyclo[2.1.1]hexane-2-carboxylateas a racemic mixture. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₉H₂₃N₂O₄:343.2; found: 342.8.

Step 2

Racemic tert-butyl3-((1,3-dioxoisoindolin-2-yl)methyl)-2-azabicyclo[2.1.1]hexane-2-carboxylate(655 mg, 1.91 mmol) was separated by chiral HPLC on a Lux Cellulose-2column using an acetronitrile eluent to afford chiral 95-A (firsteluting peak) and 95-B (second eluting peak) in enantioenriched form.For 95-A: 144 mg, 98% ee (absolute stereochemistry unknown). For 95-B:242 mg, 49% ee (absolute stereochemistry unknown).

Example 96 Preparation of Compound 96 (1R,3R,11aS)-6-hydroxy-5,7-dioxo-N-(2,4,6-trifluorobenzyl)-2,3,5,7,11,11a-hexahydro-1H-1,3-methanopyrido[1,2-a]pyrrolo[1,2-d]pyrazine-8-carboxamide

Step 1

To a solution of intermediate 95-A (141 mg, 0.41 mmol, 98% ee, unknownabsolute stereochemistry) in 9 mL ethanol was added hydrazine hydrate(0.5 mL, 10.3 mmol) and stirred at 70° C. for 18 hours to afford 96-A ofunknown absolute stereochemistry. Solids were removed by filtration andthe filtrate concentrated and carried on as crude.

Step 2

A mixture of crude 96-A (0.41 mmol assumed), 96-B (430 mg, 1.25 mmol),and sodium bicarbonate (69 mg, 0.82 mmol) in 2 mL water and 2 mL ethanolwere stirred at room temperature for 18 hours, after which the reactionmixture was diluted with water and thrice extracted to ethyl acetate.The combined organic phases were dried over magnesium sulfate, filtered,concentrated. The crude residue (222 mg) was dissolved in 1.5 mL DCM and4 N HCl in dioxane (4 mL) was added and stirred for 90 minutes at roomtemperature. The mixture was concentrated to dryness and coevaporatedwith toluene. The crude residue and DBU (0.3 mL, 2.0 mmol) in 6 mLmethanol was stirred at 50° C. for 90 minutes. The reaction mixture wasthen concentrated onto silica gel and purified by flash chromatography(0-10% MeOH/DCM) to afford 96-C. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₂N₂O₅: 395.16; found: 395.2.

Step 3

A mixture of 96-C (112 mg, 0.28 mmol), 1M aqueous potassium hydroxide (1mL), 4 mL methanol, and 4 mL THF was stirred at room temperature for 3hours, at which point the mixture was diluted with dichloromethane,acidified by addition of 1M aqueous hydrogen chloride, and the organicphase extracted to dichloromethane. The combined organics were dried,filtered, and concentrated from toluene. After drying under vacuum, theresidue was suspended in 1.5 mL DCM and trifluorobenzylamine (62 mg,0.38 mmol), HATU (220 mg, 0.58 mmol), and N,N-diisopropylethylamine(DIPEA) (0.15 mL, 0.86 mmol) were added. This reaction mixture wasstirred at room temperature for 2 hours to afford 96-D which was carriedforward as crude.

Step 4

Trifluoroacetic acid (1.7 mL, 22.2 mmol) was added to the crude reactionsolution containing 96-D from the prior step and the reaction mixtureallowed to stir at room temperature for 90 minutes. 1 mL of DMF was thenadded, the reaction mixture concentrated down to −1 mL, filtered, andpurified by preparative HPLC (ACN/water+0.1% TFA) to afford compound 96(unknown absolute stereochemistry). ¹H-NMR (400 MHz, DMSO-d₆) δ10.45-10.35 (m, 1H), 8.39 (s, 1H), 7.23-7.09 (m, 2H), 4.67 (dd, J=12.6,4.8 Hz, 2H), 4.53 (d, J=5.5 Hz, 2H), 4.20 (dd, J=11.9, 3.8 Hz, 1H),4.05-3.95 (m, 1H), 2.96-2.88 (m, 1H), 2.16 (d, J=7.0 Hz, 1H), 1.97 (d,J=7.0 Hz, 1H), 1.68-1.60 (m, 1H), 1.53-1.45 (m, 1H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₀H₁₆F₃N₃O₄: 420.12; found: 420.2.

Example 97 Preparation of Compound 97(1S,3S,11aR)-6-hydroxy-5,7-dioxo-N-(2,4,6-trifluorobenzyl)-2,3,5,7,11,11a-hexahydro-1H-1,3-methanopyrido[1,2-a]pyrrolo[1,2-d]pyrazine-8-carboxamide

Compound 97 (49% ee, unknown absolute stereochemistry) was prepared in asimilar manner to compound 96 using intermediate 95-B (49% ee, unknownabsolute stereochemistry) in place of enantiomerically oppositeintermediate 95-A. ¹H-NMR (400 MHz, DMSO-d₆) δ 10.39 (t, J=5.7 Hz, 1H),8.42 (s, 1H), 7.25-7.13 (m, 2H), 4.73-4.66 (m, 2H), 4.54 (d, J=5.7 Hz,2H), 4.20 (dd, J=12.3, 3.9 Hz, 1H), 4.01 (t, J=12.4 Hz, 1H), 2.93 (dd,J=6.7, 3.4 Hz, 1H), 2.19-2.14 (m, 1H), 1.97 (d, J=8.3 Hz, 1H), 1.65 (dd,J=10.4, 7.9 Hz, 1H), 1.49 (dd, J=10.5, 7.7 Hz, 1H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₂₀H₁₆F₃N₃O₄: 420.12; found: 420.2.

Example 98 Preparation of Compound 98(1S,4R,12aR)-3,3-difluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

98-A (0.5 g, 1.87 mmol) was dissolved in DCM (20 mL) and cooled to 0° C.under Nitrogen. Dess-Martin Periodinane (1.59 g, 3.74 mmol) was addedslowly. The mixture was stirred for 2 h at room temperature, quenchedwith Na₂S₂O₃/NaHCO₃ (7:1) aqueous saturated solution (160 mL) andstirred vigorously until two layers were separated. The crude productwas twice extracted with DCM. The combined organic layers was dried oversodium sulfate and concentrated. The crude product was purified by flashchromatography on silica gel with 0-20% MeOH/DCM to afford 98-B. ¹H-NMR(400 MHz, Chloroform-d) δ 4.34-4.05 (m, 1H), 3.97-3.75 (m, 1H), 3.69 (s,3H), 2.89 (dd, J=4.4, 2.1 Hz, 1H), 2.30-1.97 (m, 3H), 1.56 (d, J=11.3Hz, 1H), 1.35 (s, 9H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₃H₁₉NO₅:269.13; found: 270.78.

Step 2

A solution of 98-B (504 mg, 1.87 mmol) in DCM (15 mL) was stirred at 0°C. DAST (1 ml) was added drop wise to the reaction mixture. Afterovernight stirring at room temperature, the reaction mixture was cooledback to 0° C. Saturated NaHCO₃ (10 mL) was added slowly. The mixture wasextracted with twice with DCM and dried over Na₂SO₄. Afterconcentrating, the residue was purified by flash chromatography 0-50%EtOAc/hexanes to give 98-C. ¹H-NMR (400 MHz, Chloroform-d) δ 4.45-4.18(m, 1H), 3.85 (m, 1H), 3.72 (d, J=1.5 Hz, 3H), 2.72 (ddd, J=5.1, 3.2,1.6 Hz, 1H), 2.27-1.52 (m, 4H), 1.41 (d, J=21.9 Hz, 9H). ¹⁹F-NMR (376MHz, Chloroform-d) δ −91.72˜−93.99 (m), −113.65˜−115.98 (m). LCMS-ESI⁺(m/z): [M+H]⁺ calculated for C₁₃H₁₉F₂NO₄: 291.13; found: 291.55.

Step 3

98-C (476 mg, 1.634 mmol) in THF (20 mL) was stirred at 0° C. as 2.0 MLiBH₄ in THF (2.4 mL, 4.8 mmol) was added. The mixture was warmed toroom temperature and stirred for 4 h. The reaction mixture was quenchedwith ice and diluted with EtOAc and saturated NH₄Cl (some H₂ evolution).After the two phases were separated, the organic fraction was washedwith brine, dried (Na₂SO₄), and concentrated. The crude product of 98-Dwas used as is for the next step. LCMS-ESI (m/z): [M+H]⁺ calculated forC₁₂H₁₉F₂NO₃: 263.13; found: 164.10.

Step 4

98-D (1.634 mmol), phthalimide (0.36 g, 2.4 5 mmol), and PPh₃ (0.855 g,3.26 mmol) in THF (10 mL) was stirred at 0° C. bath as DIAD (0.642 mL,3.26 mmol) was added. After addition, the mixture was stirred at 0° C.for 30 min and then at room temperature for 16 h. It was diluted withEtOAc, and saturated NH₄Cl. After stirring for 5 min, a solid wasfiltered off and the two phases were separated. The organic phase waswashed with brine, dried (Na₂SO₄), and concentrated. The crude productwas purified by flash chromatography with 0-50% EA/Hex as eluents togive 98-E. ¹H-NMR suggests a mixture of two rotamers. ¹H-NMR (400 MHz,Chloroform-d) δ 7.89-7.80 (m, 2H), 7.78-7.66 (m, 2H), 5.02 (ddt, J=16.6,12.5, 6.3 Hz, 1H), 4.24 (d, J=71.8 Hz, 1H), 4.10-3.92 (m, 1H), 3.83-3.51(m, 2H), 2.46 (s, 1H), 2.21-1.98 (m, 2H), 1.87-1.62 (m, 2H), 1.31 (d,J=8.5 Hz, 9H); ¹⁹F-NMR (376 MHz, Chloroform-d) 6-91.22˜−93.58 (m),−113.20˜−115.45 (m). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₀H₂₂F₂N₂O₄: 392.15; found: 393.3.

Step 5

To a solution of 98-E (696 mg, 1.774 mmol) in EtOH (10 mL) was addedhydrazine hydrate (1 mL) at room temperature and the resulting solutionwas stirred at room temperature for 2 h. The mixture was diluted withethyl ether (30 mL) and stirred at 0° C. for 60 min before filtration.The filtrate was concentrated and the residue was dissolved in CH₂Cl₂and filtered. The filtrate was concentrated and purified by flashchromatography on silica gel with 0-20%/o MeOH (0.2% TEA)/DCM to give98-F. ¹H-NMR (400 MHz, Chloroform-d) δ 4.91 (p. J=6.2 Hz, 1H), 4.29-3.97(m, 1H), 3.36-2.93 (m, 2H), 2.49 (qt, J=8.8, 5.2 Hz, 2H), 2.08 (dddd,J=25.5, 14.0, 7.1, 4.9 Hz, 1H), 1.89-1.49 (m, 4H), 1.41 and 1.21 (d,J=6.2 Hz, 9H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −91.63˜−93.16 (m),−113.11˜−115.08 (m). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₂H₂₀F₂N₂O₂: 262.15; found: 262.8.

Step 6, 7 and 8

The mixture of 98-G (375.8 mg, 1.55 mmol), 98-E (370 mg, 1.41 mmol), andNaHCO₃ (261 mg, 3.10 mmol) in water (5 mL) and EtOH (5 mL) was stirredat room temperature for 2 h. The mixture was diluted with brine andextracted with EtOAc (×2). The extracts were combined, dried (Na₂SO₄),concentrated, and dried in vacuo to afford crude A. LCMS-ESI⁺ (m/z):[M+H]⁺ 591.59. Crude A (1.38 mmol) in CH₂Cl₂ (5 mL) was added 4 N HCl indioxane (5 mL). After 2 h at room temperature, mixture was concentratedto dryness. It was co-evaporated with toluene once and dried in vacuo toafford crude B. B (1.38 mmol+0.442 mmol) and DBU (3 mL, 11 mmol) inanhydrous MeOH (15 mL) were stirred at 50° C. bath for 40 min. Themixture was concentrated. The residue was purified by flashchromatography (80 g column) using 0-20% MeOH/DCM as eluents to give98-H. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₃H₂₂F₂N₂O₅: 444.15;found: 445.36 (90%), 431.18 (10%).

Steps 9, 10 and 11

The remaining steps were performed using procedures similar to Example41 to afford desired compound 98. ¹H-NMR (400 MHz, Chloroform-d) δ 10.29(d, J=6.1 Hz, 1H), 8.34 (s, 1H), 6.65 (dd, J=8.7, 7.5 Hz, 2H), 4.83 (s,1H), 4.72-4.58 (m, 2H), 4.36-4.10 (m, 2H), 4.05 (t, J=11.5 Hz, 1H), 2.97(d, J=4.4 Hz, 1H), 2.49-2.08 (m, 3H), 2.12-1.94 (m, 1H). ¹⁹F-NMR (376MHz, Chloroform-d) δ −92.08-−93.57 (m, 1F), −108.92 (ddd, J=15.1, 8.8,6.3 Hz, 1F), −109.30˜−110.65 (m, 1F), −112.16 (p, J=7.3 Hz, 2F).LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₂₁H₁₆F₅N₃O₄: 469.11; found:470.23.

Example 99 Preparation of Compound 99(1R,3S,4R,12aR)-7-hydroxy-3-methyl-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

To a stirred solution of 99-A (1 g, 3.71 mmol) in THF (20 mL) was addeddropwise a solution of the Tebbe reagent (0.5 M in toluene, 14.85 mL,7.42 mmol) at 0° C. After addition, the brown solution was allowed towarm to room temperature slowly and was stirred at room temperature for2 h. The reaction was quenched carefully by the addition of saturatedNaHCO₃ solution at 0° C., and the mixture was stirred at roomtemperature for 15 minutes. The mixture was filtered through celite, andthe filter cake was washed with ether and DCM (1:1) twice. Afterseparated layers, the organics were combined and concentrated in vacuo,and the residue was purified by column chromatography on silica gelcolumn with 0-50% EtOAc/hexanes to afford 99-B. ¹H-NMR (400 MHz,Chloroform-d) δ 5.06 (dt, J=48.6, 2.6 Hz, 1H), 4.73 (d, J=7.0 Hz, 1H),4.42 (d, J=61.8 Hz, 1H), 3.81 (d, J=48.2 Hz, 1H), 3.73 (d, J=1.6 Hz,3H), 2.74 (dd, J=9.4, 4.4 Hz, 1H), 2.38 (ddt, J=13.5, 4.5, 2.5 Hz, 1H),2.18-2.06 (m, 1H), 1.99 (dt, J=10.2, 2.4 Hz, 1H), 1.58 (s, 1H), 1.42 (d,J=25.5 Hz, 9H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₄H₂₁NO₄:267.15; found: 267.65.

Step 2

A mixture of 99-B (675 mg, 2.506 mmol) and 20% Pd(OH)₂/C (500 mg) inEtOH (50 mL) was stirred under H₂ atmosphere. The mixture was filteredthrough Celite and the filtrate was concentrated to give 99-C. ¹H-NMR(400 MHz, Chloroform-d) δ 4.23-3.99 (m, 1H), 3.77-3.64 (m, 4H), 2.55 (d,J=4.8 Hz, 1H), 2.14-1.86 (m, 3H), 1.42 (d, J=24.2 Hz, 9H), 0.96 (d,J=6.6 Hz, 3H), 0.85 (ddd, J=12.5, 4.8, 2.4 Hz, 1H). LCMS-ESI⁺ (m/z):[M+H]⁺ calculated for C₁₄H₂₃NO₄: 269.16; found: 269.69.

Step 3

99-C (670 mg, 2.488 mmol) in THF (20 mL) was stirred at 0° C. as 2.0 MLiBH₄ in THF (3.7 mL, 7.46 mmol) was added. The mixture was warmed toroom temperature and stirred for 4 h. The reaction mixture was quenchedwith ice and diluted with EtOAc and saturated NH₄Cl (some H₂ evolution).After two phases were separated, the organic fraction was washed withbrine, dried (Na₂SO₄), and concentrated. The crude alcohol 99-D was usedas is for the next step. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₃H₂₃NO₃: 241.17; found: 241.76.

Steps 4 and 5

Steps 4 and 5 were performed using procedures similar to those inExample 41 to afford 99-F. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₃H₂₄N₂O₂: 240.18; found: 241.2.

Step 6, 7 and 8

Steps 6, 7 and 8 were performed using procedures similar to that ofExample 41 to give 99-G. LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₄H₂₆N₂O₅: 422.18; found: 423.21.

Step 9, 10 and 11

The remaining steps were performed using procedures similar to Example41 to afford compound 99. ¹H-NMR (400 MHz, Chloroform-d) δ 11.71 (s,1H), 10.36 (t, J=5.7 Hz, 1H), 8.28 (s, 1H), 6.63 (t, J=8.1 Hz, 2H), 4.63(t, J=5.4 Hz, 3H), 4.12 (dd, J=12.3, 3.5 Hz, 1H), 3.83 (t, J=12.3 Hz,1H), 3.67 (dd, J=12.3, 3.4 Hz, 1H), 2.64-2.52 (m, 1H), 2.30 (ddq,J=10.5, 7.2, 3.6 Hz, 1H), 2.13 (td, J=12.1, 4.4 Hz, 1H), 1.82-1.63 (m,2H), 1.24 (d, J=3.3 Hz, 1H), 1.04 (d, J=6.9 Hz, 4H), 0.90-0.79 (m, 1H).¹⁹F-NMR (376 MHz, Chloroform-d) δ −109.20 (ddd, J=15.0, 8.8, 6.2 Hz),−112.03 (t, J=7.0 Hz). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₂₂H₂₀F₃N₃O₄: 447.14; found: 448.32.

Example 100 Preparation of Compound 100(1R,4R,12aS)—N-(2,4-difluorobenzyl)-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-ethanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL rbf was charged with 100-A (2.0 g, 7.8 mmol) in THF (20 mL).The reaction mixture was cooled to 0° C. Borane dimethyl sulfide (2 N inTHF, 17.6 mL) was slowly added in. Then the reaction mixture was stirredat room temperature for overnight. The reaction mixture was cooled backto 0° C. Methanol (8 mL) was added drop wise to quench the reaction.After concentration, the residue was purified by Combi Flash (40 gcolumn, cartridge used) using hexanes-EA as eluents to afford 100-B.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 242.

Step 2

A 100-mL rbf was charged with 100-B (1.8 g, 7.4 mmol),triphenylphosphine (4.3 g, 16.2 mmol) and phthalimide (1.8 g, 12.2 mmol)in THF (30 mL). Then the reaction mixture was cooled to 0° C. withstirring. DIAD (3.2 mL, 16.2 mmol) was slowly added to the reactionmixture. The reaction mixture was stirred at room temperature forovernight. After concentration, the residue was purified by Combi Flash(80 g column, cartridge used) using hexanes-EA as eluents to afford100-C. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 371.

Step 3

To a solution of 100-C (2.5 g, 6.8 mmol) in EtOH (50 mL) was addedhydrazine monohydrate (1.7 mL). The reaction mixture was heated to 70°C. with stirring for 3 hours. After filtration to remove the solid, thefiltrate was concentrated to afford 100-D. LCMS-ESI⁺ (m/z): [M+H]⁺found: 241.

Step 4

A 100-mL rbf was charged with 100-D (1.6 g, 6.7 mmol) and 100-E (2.3 g,6.7 mmol) in ethanol (30 mL). Sodium bicarbonate (1.2 g, 1.4 mmol) inwater (30 mL) was added to the reaction mixture. Then the reactionmixture was stirred at room temperature for overnight. The mixture wasdiluted with EA (200 mL) and washed with water (2×). The aqueousfractions were extracted with EA (1×), and the organic fractions werecombined, dried (Na₂SO₄), and concentrated. The crude 100-F was used fornext step without further purification. LCMS-ESI⁺ (m/z): [M+H]⁺ found:569.

Step 5

A 100-mL rbf was charged with 100-F (3.7 g, 6.5 mmol) in 4 N HCl/dioxane (38 mL). Then the reaction mixture was stirred at roomtemperature for 1 hour. After concentration, 3.2 g intermediate wasobtained. The intermediate and DBU (5.1 g, 33.8 mmol) were dissolved intoluene (100 mL). The reaction mixture was heated to 110° C. withstirring for 1 hour. After concentration, the residue was purified byCombi Flash (80 g column, cartridge used) using hexanes-EA as eluents toafford 100-G. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 423.

Step 6

A 100-mL rbf was charged with 100-G (2.0 g, 4.7 mmol) in THF (20 mL) andMeOH (20 mL). 1 N KOH (18.9 mL) was added to the reaction mixture. Thenthe reaction mixture was stirred at room temperature for 1 hour. Thereaction mixture was acidified by adding 1 N HCl (18.9 mL). Afterconcentration, the residue was co-evaporated with toluene (3×). Thecrude acid (0.28 g, 0.72 mmol), 2, 4-difluobenzylamine (0.2 g, 1.44mmol), N,N-diisopropylethylamine (DIPEA) (0.47 g, 3.6 mmol) and HATU(0.55 g, 1.44 mmol) were dissolved in DCM (20 mL). The reaction mixturewas stirred at room temperature for 2 hours. The mixture was dilutedwith EA (100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl(2×) and dried over Na₂SO₄. After concentration, the crude was purifiedby column chromatography on silica gel with hexane-EtOAc to afford100-H. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 520.

Step 7

A 50-mL rbf was charged with 100-H (0.36 g, 0.69 mmol) in TFA (5 mL).The reaction mixture was stirred at room temperature for 30 minutes.After concentration, the crude was purified by column chromatography onsilica gel with EtOAc-MeOH to afford compound 100. ¹H-NMR (400 MHz,Chloroform-d) δ 12.25 (m, 1H), 10.47 (t, J=5.9 Hz, 1H), 8.30 (s, 1H),7.58-7.29 (m, 1H), 6.98-6.50 (m, 2H), 4.62 (dd, J=14.8, 4.9 Hz, 3H),4.22 (t, J=12.2 Hz, 1H), 4.14-4.07 (m, 1H), 3.96 (dd, J=12.2, 3.1 Hz,1H), 2.26-1.44 (m, 9H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −112.38 (t,J=7.7 Hz), −114.78 (q, J=8.5 Hz). LCMS-ESI⁺ (m/z): found: 430.

Example 101 Preparation of Compound 101(1R,4R,12aS)-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-ethanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL rbf was charged with 101-A (0.3 g, 0.72 mmol) in THF (2 mL) andMeOH (2 mL). 1 N KOH (2.1 mL) was added to the reaction mixture. Thenthe reaction mixture was stirred at room temperature for 1 hour. Thereaction mixture was acidified by adding 1 N HCl (2.1 mL). Afterconcentration, the residue was co-evaporated with toluene (3×). Thecrude acid (0.72 mmol), 2, 4, 6-trifluobenzylamine (0.23 g, 1.44 mmol),N,N-diisopropylethylamine (DIPEA) (0.47 g, 3.6 mmol) and HATU (0.55 g,1.44 mmol) were dissolved in DCM (20 mL). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted with EA(100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl (2×) anddried over Na₂SO₄. After concentration, the crude was purified by columnchromatography on silica gel with hexane-EtOAc to afford 101-B.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 538.

Step 2

A 50-mL rbf was charged with 101-B (0.36 g, 0.67 mmol) in TFA (5 mL).The reaction mixture was stirred at room temperature for 30 minutes.After concentration, the crude was purified by column chromatography onsilica gel with EtOAc-MeOH to afford compound 101. ¹H-NMR (400 MHz,Chloroform-d) δ 12.11 (s, 1H), 10.40 (t, J=5.8 Hz, 1H), 8.28 (s, 1H),6.91-6.39 (m, 2H), 4.62 (ddd, J=25.0, 6.5, 2.8 Hz, 3H), 4.21 (t, J=12.2Hz, 1H), 4.09 (dd, J=12.5, 3.0 Hz, 1H), 3.93 (dd, J=12.2, 3.1 Hz, 1H),2.35-1.39 (m, 9H). ¹⁹F NMR (376 MHz, Chloroform-d) δ −112.38 (t, J=7.7Hz), −114.78 (q, J=8.5 Hz). LCMS-ESI⁺ (m/z): found: 448.

Example 102 Preparation of Compound 102(1S,4S,12aR)-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-ethanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A 100-mL rbf was charged with 102-A (2.0 g, 7.8 mmol) in THF (20 mL).The reaction mixture was cooled to 0° C. Borane dimethyl sulfide (2 N inTHF, 17.6 mL) was slowly added in. Then the reaction mixture was stirredat room temperature for overnight. The reaction mixture was cooled backto 0° C. Methanol (8 mL) was added drop wise to quench the reaction.After concentration, the residue was purified by Combi Flash (40 gcolumn, cartridge used) using hexanes-EA as eluents to afford 102-B.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 242.

Step 2

A 100-mL rbf was charged with 102-B (1.8 g, 7.4 mmol),triphenylphosphine (4.3 g, 16.2 mmol) and phthalimide (1.8 g, 12.2 mmol)in THF (30 mL). Then the reaction mixture was cooled to 0° C. withstirring. DIAD (3.2 mL, 16.2 mmol) was slowly added to the reactionmixture. The reaction mixture was stirred at room temperature forovernight. After concentration, the residue was purified by Combi Flash(80 g column, cartridge used) using hexanes-EA as eluents to afford102-C. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 371.

Step 3

To a solution of 102-C (2.5 g, 6.8 mmol) in EtOH (50 mL) was addedhydrazine monohydrate (1.7 mL). The reaction mixture was heated to 70°C. with stirring for 3 hours. After filtration to remove the solid, thefiltrate was concentrated to afford 102-D. LCMS-ESI⁺ (m/z): [M+H]⁺found: 241.

Step 4

A 100-mL rbf was charged with 102-D (1.6 g, 6.7 mmol) and 102-E (2.3 g,6.7 mmol) in ethanol (30 mL). Sodium bicarbonate (1.2 g, 1.4 mmol) inwater (30 mL) was added to the reaction mixture. Then the reactionmixture was stirred at room temperature for overnight. The mixture wasdiluted with EA (200 mL) and washed with water (2×). The aqueousfractions were extracted with EA (1×), and the organic fractions werecombined, dried (Na₂SO₄), and concentrated. The crude 102-F was used fornext step without further purification. LCMS-ESI⁺ (m/z): [M+H]⁺ found:569.

Step 5

A 100-mL rbf was charged with 102-F (3.7 g, 6.5 mmol) in 4 N HCl/dioxane (38 mL). Then the reaction mixture was stirred at roomtemperature for 1 hour. After concentration, 3.2 g intermediate wasobtained. The intermediate and DBU (5.1 g, 33.8 mmol) were dissolved intoluene (100 mL). The reaction mixture was heated to 110° C. withstirring for 1 hour. After concentration, the residue was purified byCombi Flash (80 g column, cartridge used) using hexanes-EA as eluents toafford 102-G. LCMS-ESI⁺ (m/z): [M+H]⁺ found: 423.

Step 6

A 100-mL rbf was charged with 102-G (0.3 g, 0.72 mmol) in THF (2 mL) andMeOH (2 mL). 1 N KOH (2.1 mL) was added to the reaction mixture. Thenthe reaction mixture was stirred at room temperature for 1 hour. Thereaction mixture was acidified by adding 1 N HCl (2.1 mL). Afterconcentration, the residue was co-evaporated with toluene (3×). Thecrude acid (0.72 mmol), 2, 4, 6-trifluobenzylamine (0.23 g, 1.44 mmol),N,N-diisopropylethylamine (DIPEA) (0.47 g, 3.6 mmol) and HATU (0.55 g,1.44 mmol) were dissolved in DCM (20 mL). The reaction mixture wasstirred at room temperature for 2 hours. The mixture was diluted with EA(100 mL) and washed with saturated NaHCO₃ (2×), saturated NH₄Cl (2×) anddried over Na₂SO₄. After concentration, the crude was purified by columnchromatography on silica gel with hexane-EtOAc to afford 102-H.LCMS-ESI⁺ (m/z): [M+H]⁺ found: 538.

Step 7

A 50-mL rbf was charged with 102-H (0.36 g, 0.67 mmol) in TFA (5 mL).The reaction mixture was stirred at room temperature for 30 minutes.After concentration, the crude was purified by column chromatography onsilica gel with EtOAc-MeOH to afford compound 102. ¹H-NMR (400 MHz,Chloroform-d) δ 12.13 (s, 1H), 10.40 (t, J=5.8 Hz, 1H), 8.28 (s, 1H),6.64 (t, J=8.1 Hz, 2H), 4.89-4.41 (m, 3H), 4.22 (t, J=12.2 Hz, 1H), 4.09(dd, J=12.3, 3.1 Hz, 1H), 3.95 (dd, J=12.1, 4.1 Hz, 1H), 2.45-1.60 (m,9H). ¹⁹F-NMR (376 MHz, Chloroform-d) δ −109.26 (ddd, J=15.1, 8.8, 6.3Hz), −111.99 (t, J=6.9 Hz). LCMS-ESI⁺ (m/z): found: 448.

Example 103 Preparation of Compound 103(1R,4R,12aR)-2,3-difluoro-7-hydroxy-6,8-dioxo-N-(2,4,6-trifluorobenzyl)-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide

Step 1

A solution of (1R,3R,4R,5R,6S)-methyl5,6-dihydroxy-2-((S)-1-phenylethyl)-2-azabicyclo[2.2.1]heptane-3-carboxylate(2.0 g, 6.9 mmol) in DCM (27 mL) was cooled to −78° C. in a dryice/acetone bath. To this solution was added DAST (2.18 ml, 16.48 mmol)via plastic tipped pipette. The solution was stirred at −78° C. for 30minutes after which time it was removed from the bath, let warm slowlyto room temperature, and stirred at room temperature for one hour. Thereaction was quenched by slow addition of the reaction mixture to astirring solution of saturated sodium bicarbonate (150 mL) via plastictipped pipette. The layers were separated and the aqueous layer wasback-extracted with dichloromethane. The combined organic layers weredried over magnesium sulfate, filtered and concentrated in vacuo. Thecrude product was purified by silica gel chromatography (7-28% ethylacetate/hexane) to provide 103-A. ¹H-NMR (400 MHz, Chloroform-d) δ7.43-7.16 (m, 5H), 5.01-4.60 (m, 2H), 3.85 (q, J=7.1, 6.6 Hz, 1H), 3.55(s, 2H), 3.53-3.42 (m, 2H), 2.76 (dq, J=5.1, 2.0 Hz, 1H), 2.19-2.07 (m,1H), 2.03-1.88 (m, 1H), 1.39 (d, J=6.7 Hz, 3H).

Steps 2 and 3

To a solution of 103-A (0.96 g, 3.24 mmol) in Ethanol (36.01 ml) and1.25M HCl-ethanol (4.09 ml) was added 20% PdOH/C (1.14 g, 1.62 mmol) thesuspension was stirred under an atmosphere of hydrogen for 22 hours.After filtering through Celite, the cake was washed with EtOH, thefiltrate was concentrated under vacuum to dryness to afford the crudedeprotected product which was assumed to be 3.24 mmol for next step.LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₈H₁₂F₂NO₂: 192.08; found:192.110.

To the crude residue (0.62 g, 3.24 mmol) and Di-tert-butyl dicarbonate(1.06 g, 4.86 mmol) in 2-Methyltetrahydrofuran (32.43 ml) was addedN,N-diisopropylethylamine (0.56 ml, 0 mol). Upon completion, thereaction mixture was diluted with water, extracted into EtOAC (2×) andthe organic fractions were washed with water, combined, dried (Na₂SO₄),and concentrated. The residue was purified by silica columnchromatography (0-55% EtOAc/Hexanes) to afford 103-B. ¹H-NMR (400 MHz,Chloroform-d) δ 5.12-5.01 (m, 1H), 4.92 (s, 1H), 4.49 (s, 1H), 4.14 (d,J=14.7 Hz, 1H), 3.75 (s, 3H), 2.91 (s, 1H), 2.24-1.98 (m, 2H), 1.47 (s,5H), 1.38 (s, 5H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated for C₁₃H₂₀F₂NO₄:292.13; found: 291.75.

Step 4

A solution of 103-B (0.68 g, 2.33 mmol) in THF (15 ml) was stirred in anice bath as 1.0 M LiBH4 in THF (4.65 ml) was added and the resultingmixture was stirred at 0° C. for 30 minutes at which time it was shownto be complete by TLC. The reaction mixture was carefully treated withwater (0.3 mL), then with NaOH (˜15%, 3.5M, 0.3 mL), then finally withadditional water (0.9 mL). The mixture was stirred at room temperaturefor 15 minutes, and the ppt that formed was filtered, washed withdiethyl ether and the supernate was concentrated to afford 103-C. ¹H-NMR(400 MHz, Chloroform-d) δ 4.83 (s, 1H), 4.56 (d, J=10.5 Hz, 1H), 4.37(s, 1H), 3.78-3.47 (m, 3H), 2.76 (s, 1H), 2.36-2.18 (m, 1H), 2.17-1.98(m, 1H), 1.55 (s, 1H), 1.48 (s, 9H).

Steps 5 and 6

A mixture of 103-C (0.59 g, 2.25 mmol), phthalimide (0.53 g, 3.6 mmol)and triphenylphosphine (1.3 g, 4.95 mmol) in THF (11 ml) was cooled inan ice bath. Diisopropyl Azodicarboxylate (0.97 ml, 4.95 mmol) wasadded. The mixture was then warmed up to room temperature and stirredfor 14 h and then concentrated in vacuo. The residue was dissolved inether, stirred for 1 h, then the solids were filtered off and thefiltrate was concentrated. The residue was purified by silica columnchromatography (10-31-91% EtOAc/hexanes) to afford the protected aminocompound (assumed 2.25 mmol of product). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₀H₂₃F₂N₂O₄: 393.15; found: 392.77.

A solution of the protected amino compound (0.88 g, 2.25 mmol) andhydrazine hydrate (0.46 ml, 9.52 mmol) in ethanol (22 ml) was stirred at60° C. for 2 h. The reaction mixture was cooled in an ice bath, ether(10 ml) was added and the mixture was stirred for 30 min. The solidformed was filtered off and the filtrate was concentrated under vacuumto dryness to give 103-D. ¹H-NMR (400 MHz, Chloroform-d) δ 5.17-4.61 (m,2H), 4.37 (s, 1H), 3.80 (s, 1H), 3.11-2.77 (m, 1H), 2.01 (s, 2H), 1.87(s, 1H), 1.83 (d, J=7.4 Hz, 1H), 1.46 (s, 9H), 1.30 (d, J=6.4 Hz, 1H),1.27 (d, J=6.3 Hz, 3H). LCMS-ESI⁺ (m/z): [M+H]⁺ calculated forC₁₂H₂₀F₂N₂O₂: 263.15; found: 262.86.

Steps 7, 8 and 9

Compound 103 was prepared in a similar manner to compound 60 using 103-Din place of 41-E and using (2,4,6-trifluorophenyl)methanamine in placeof (2,3-dichlorophenyl)methanamine. A single diastereomer resulted. Thestereochemistry of the fluorines is unknown. ¹H-NMR (400 MHz,Chloroform-d) δ 8.08 (s, 1H), 6.46-6.27 (m, 2H), 4.95 (d, J=53.5 Hz,1H), 4.65 (d, J=54.9 Hz, 1H), 4.45 (s, 1H), 4.33 (d, J=5.6 Hz, 2H), 3.84(t, J=3.6 Hz, 2H), 2.75 (s, 1H), 2.28 (p, J=1.9 Hz, 2H), 2.20 (s, 1H),1.91 (dd, J=33.3, 15.2 Hz, 1H), 0.95 (s, 1H). LCMS-ESI⁺ (m/z): [M+H]⁺calculated for C₂₁H₁₇F₅N₃O₄: 470.11; found: 0.470.13.

Antiviral Assay Example 104 Antiviral Assays in MT4 Cells

For the antiviral assay utilizing MT4 cells, 0.4 μL of 189X testconcentration of 3-fold serially diluted compound in DMSO was added to40 μL of cell growth medium (RPMI 1640, 10% FBS, 1%penicilline/Streptomycine, 1% L-Glutamine, 1% HEPES) in each well of384-well assay plates (10 concentrations) in quidruplicate.

1 mL aliquots of 2×10⁶ MT4 cells are pre-infected for 1 and 3 hoursrespectively at 37° C. with 25 μL (MT4) or of either cell growth medium(mock-infected) or a fresh 1:250 dilution of an HIV-IIIb concentratedABI stock (0.004 m.o.i. for MT4 cells). Infected and uninfected cellsare diluted in cell growth medium and 35 μL of 2000 (for MT4) cells isadded to each well of the assay plates.

Assay plates were then incubated in a 37° C. incubator. After 5 days ofincubation, 25 μL of 2× concentrated CellTiter-Glo™ Reagent (catalog #G7573, Promega Biosciences, Inc., Madison, Wis.) was added to each wellof the assay plate. Cell lysis was carried out by incubating at roomtemperature for 2-3 minutes, and then chemiluminescence was read usingthe Envision reader (PerkinElmer).

Compounds of the present invention demonstrate antiviral activity inthis assay as depicted in Table 1 below. Accordingly, the compounds ofthe invention may be useful for treating the proliferation of the HIVvirus, treating AIDS, or delaying the onset of AIDS or ARC symptoms.

TABLE 1 nM in MT-4 Compound Number EC₅₀ CC₅₀ 1 2.6 5819 2 2.2 3111 3 2.038446 4 14.8 45769 5 8.1 10452 6 5.3 53192 7 3.5 15610 8 2.5 13948 9 5.113451 10 6.1 3670 11 4.9 10274 12 5.9 3337 13 46.0 12666 14 65.5 4939 152.2 16268 16 1.5 13633 17 5.9 6613 18 4.1 10263 19 2.8 38690 20 3.327990 21 38.3 13010 22 64.3 4433 23 2.3 13444 24 6.1 12074 25 26.2 523326 10.3 8836 27 4.4 8751 28 15.6 18687 29 13.9 9446 30 4.0 6828 31 9.04525 32 14.0 4684 33 43.5 3971 34 422.1 3585 35 157.0 15546 36 7.6 1142437 10.2 19486 38 1.7 10223 39 3.6 12174 40 2.4 9560 41 2.1 15675 42 2.53544 43 6.9 10321 44 2.3 9869 45 2.4 15765 46 2.6 19295 47 1.9 11301 482.7 13967 49 33.3 52219 50/51 1.9 37173 (racemic mixture) 52 15.0 1294353 14.3 3347 54 15.6 3236 55 1.5 11100 56 3.1 17238 57 2.3 11751 58 1.57694 59 3.1 22200 60 2.1 3308 61 1.8 25881 62 9.2 3492 63 2.5 3164 643.5 3332 65 2.4 2508 66 9.4 11848 67 10.7 2981 68 2.7 4175 69 1.9 476770 5.1 8413 71 2.6 4660 72 4.3 6255 73 1.8 9194 74 29.3 4340 75 2.8 529276 17.8 34581 77 5.6 10145 78 5.6 3198 79 3.4 12092 80 4.6 5045 81 1.912298 82 2.9 30434 83 1.9 27501 84 2.9 9727 85 2.0 10378 86 2.3 22405 882.9 3230 89 8.4 4629 90 5.7 8086 91 5.0 7183 92 18.6 4553 93 2.2 6158 9411.5 51173 96 2.6 26586 97 2.1 17341 98 2.4 17947 99 2.0 8475 100 2.211580 101 2.1 11585 102 2.3 12042 103 10.3 35127

Example 105 Human PXR Activation Assay

Luciferase Reporter Gene Assay.

A stably transformed tumor cell line (DPX2) was plated on 96-wellmicrotiter plates. DPX2 cells harbor the human PXR gene (NR1I2) and aluciferase reporter gene linked to two promoters identified in the humanCYP3A4 gene, namely XREM and PXRE. The cells were treated with sixconcentrations of each compound (0.15˜50 μM) and incubated for 24 hours.The number of viable cells was determined and the reporter gene activitywas assessed. Positive control: Rifampicin at 6 concentrations (0.1˜20μM). % E_(max) relative to the maximum fold induction by 10 or 20 μM RIFwas calculated for test compounds according to the following equationwhich adjusts for the DMSO background: % E_(max)=(Foldinduction−1)/(Maximum fold induction by RIF−1)×100%.

TABLE 2 Compound % E_(max) at Number 15 μM 2 4.5 3 7.5 4 3 5 32 6 0 7 68 7 9 7 10 19 15 20 16 17 17 7 18 4 19 2 20 2 23 45 28 6 29 3 32 14 3317 36 3 37 2 38 7 39 6 40 0 41 11.5 42 21 43 18 44 4 45 19 46 34 47 1148 5 54 2 55 24 56 3 57 3 58 1 59 4 60 3 61 1 63 13 64 8 66 0 67 0 68 669 5 70 10 71 3 72 4 73 7 75 0 77 11 79 0 80 2 81 1 82 1 83 1 84 21 8577 86 30 88 27 89 5 90 11 91 3 92 3 93 9 96 11 97 9 98 0 99 17 100 45102 123 103 0

Example 106 OCT2 Inhibition Assay

The dose dependent inhibition of OCT2 mediated uptake of a modelsubstrate ¹⁴C-Tetraethylammonium (TEA) by test compounds was studied inwild-type and OCT2-transfected MDCKII cells at 7 concentrations from0.014 μM to 10 μM.

MDCKII cells were maintained in minimal essential medium (MEM) with 1%Pen/Strep, 10% fetal bovine serum, and 0.25 mg/mL hygromycin B in anincubator set at 37° C., 90% humidity and 5% CO₂. 24 hours prior toassay, media containing 5 mM sodium butyrate were added to MDCKII cellsin flasks, and cells were grown to 80-90% confluence. On assay day,cells were trypsinized and resuspended in Krebs-Henseleit Buffer (KHB),pH 7.4 at 5×10⁶ million cells/mL. Cells were preincubated for 15 min inassay plate before addition of test compound or substrate.

Test compounds were serially diluted in DMSO and then spiked (2 μL) intoin 0.4 mL KHB buffer containing wild-type or OCT2-transfected cells andincubated for 10 minutes. Assay was initiated with the addition of 0.1mL of 100 μM ¹⁴C-TEA in KHB buffer (20 μM final concentration aftermixing). The concentration of TEA is based on the K_(m). After 10minutes of incubation, the assay mixture was quenched with addition of0.5 mL of ice-cold IX PBS buffer. Samples were then centrifuged at 1000rpm for 5 min and supernatants were removed. Wash steps were repeatedfour times with ice-cold PBS. Finally, the cell pellets were lysed with0.2N NaOH and let sit at room temperature for at least 30 min to ensurecomplete lysis.

Samples were then counted on liquid scintillation counter and dpm countswere used to perform the following calculations. The % inhibition wascalculated as follows: %inhibition=[1−{[OCT2]_(i)−[WT]_(ni)}/{[OCT2]_(ni)−[WT]_(ni)}]*100 where,[OCT2]_(i) represents the dpm count in the presence of test compound foreither OCT2 cells, [OCT2]_(ni) represents the dpm count in the absenceof test compound for OCT2 cells and [WT]_(ni) represents the dpm countin the absence of test compound for wild type cells, respectively.

TABLE 3 Compound Number IC₅₀ (nM) 2 240 3 250 5 2230 11 10000 13 610 3610000 39 358 40 204 41 2823 42 487 45 137 47 6200 48 4909 55 476 63 4264 94 77 3830 82 10000 83 10000 96 1357 98 3726 99 1506 100 450

The data in Tables 1, 2 and 3 represent an average over time of eachassays for each compound. For certain compounds, multiple assays havebeen conducted over the life of the project. Thus, the data reported inTables 1, 2 and 3 include the data reported in the priority documents,as well as data from assays run in the intervening period.

Example 107 Pharmacokinetic Analysis Following Oral or IntravenousAdministration to Beagle Dogs

Pharmacokinetic analysis was performed on various test compoundsfollowing intravenous or oral administration to beagle dogs.

For pharmacokinetic analysis of intravenously administered compounds,the test compounds were formulated in 5% Ethanol, 55% PEG 300, and 40%water at 0.1 mg/mL for IV infusion. For pharmacokinetic analysis oforally administered compounds, the test compounds were formulated as anaqueous suspension in 0.1% Tween 20, 0.5% HPMC LV100 in Di Water at 1mg/kg.

Each dosing group consisted of 3 male, non-naïve purebred beagle dogs.At dosing, the animals weighed between 10 to 13 kg. The animals werefasted overnight prior to dose administration and up to 4 hr afterdosing. For studies of intravenous administration, the test article wasadministered to the animals by intravenous infusion over 30 min. Therate of infusion was adjusted according to the body weight of eachanimal to deliver a dose of 0.5 mg/kg. For studies of oraladministration, the test article was administered according to the bodyweight of each animal to deliver a dose of 1 mg/kg.

For pharmacokinetic analysis of intravenously administered compounds,serial venous blood samples (approximately 1 mL each) were taken fromeach animal at 0, 0.250, 0.483, 0.583, 0.750, 1.00, 1.50, 2.00, 4.00,8.00, 12.0, and 24.0 hours after dosing. The blood samples werecollected into Vacutainer™ tubes containing EDTA-K2 as theanti-coagulant and were immediately placed on wet ice pendingcentrifugation for plasma. An LC/MS/MS method was used to measure theconcentration of the test compound in plasma. An aliquot of 100 μL ofeach plasma sample was added to a clean 96 well plate, and 400 μL ofcold acetonitrile/internal standard solution (ACN)/(ISTD) was added.After protein precipitation, an aliquot of 110 μL of the supernatant wastransferred to a clean 96-well plate and diluted with 300 μL of water.An aliquot of 25 μL of the above solution was injected into a TSQQuantum Ultra LC/MS/MS system utilizing a Hypersil Gold C₁₈ HPLC column(50×3.0 mm, 5 μm; Thermo-Hypersil Part #25105-053030). An Agilent 1200series binary pump (P/N G1312A Bin Pump) was used for elution andseparation, and an HTS Pal autosampler (LEAP Technologies, Carrboro,N.C.) was used for sample injection. A TSQ Quantum Ultra triplequadrupole mass spectrometer was utilized in selective reactionmonitoring mode (Thermo Finnigan, San Jose, Calif.). Liquidchromatography was performed using two mobile phases: mobile phase Acontained 1% acetonitrile in 2.5 mM ammonium formate aqueous solutionwith pH of 3.0, and mobile phase B contained 90% acetonitrile in 10 mMammonium formate with pH of 4.6. Non-compartmental pharmacokineticanalysis was performed on the plasma concentration-time data. Theresulting data are shown in the first three columns of Table 4. In Table4, CL refers to clearance, which characterizes the rate at which drug isremoved from plasma. The lower the clearance of a drug is, the longerthe elimination half-life is in the body. V_(ss) refers to the steadystate volume of distribution and indicates how well a drug isdistributed into the tissues. The larger the V_(ss) is, the longer theelimination half-life is in the body. MRT refers to mean residence time,which is a measure of the average time molecules exist in the body.

For pharmacokinetic analysis of orally administered compounds, serialvenous blood samples (approximately 0.3 mL each) were taken from eachanimal at time points of 0, 0.25, 0.50, 1.0, 2.0, 4.0, 6.0, 8.0, 12.0and 24.0 hours after dosing. Blood samples were collected, prepared andanalyzed in a similar way to the intranveous studies described above.Non-compartmental pharmacokinetic analysis was performed on the plasmaconcentration-time data. The resulting data are shown in the last threecolumns of Table 4. In Table 4, F (%) refers to oral bioavailability.C_(max) refers to the peak plasma concentration of the compound afteradministration. AUC refers to area under the curve and is a measure oftotal plasma exposure of the indicated compound.

TABLE 4 F (%) AUC aqueous C_(max) (μM) (μM * h) Compound CL V_(ss) MRTsus- aqueous aqueous # (L/h/kg) (L/kg) (h) pension suspension suspension98 0.047 0.16 3.3 n/a n/a n/a 83 0.161 0.38 2.4 n/a n/a n/a 55 0.0580.24 4.2 n/a n/a n/a 77 0.300 0.64 2.2 n/a n/a n/a 41 0.015 0.11 7.510.7 2.4 16.3 42 0.020 0.15 7.1 28.0 4.5 28.6 47 0.014 0.10 7.4 12.6 2.820.4 8 0.498 0.87 1.8 n/a n/a n/a 7 0.510 1.20 2.3 n/a n/a n/a 3 0.0470.23 4.9 18.7 1.2  9.2 2 0.030 0.20 6.5 40.7 7.8 66.1

All of the U.S. patents, U.S. patent application publications, U.S.patent applications, foreign patents, foreign patent applications andnon-patent publications referred to in this specification areincorporated herein by reference, in their entirety to the extent notinconsistent with the present description.

From the foregoing it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention. Accordingly, the invention is notlimited except as by the appended claims.

1. A compound having the following Formula (I):

or a stereoisomer or pharmaceutically acceptable salt thereof, wherein:X is —O— or —NZ³— or —CHZ³—; W is —CHZ²—; Z¹, Z² and Z³ are each,independently, hydrogen or C₁₋₃alkyl, or wherein Z¹ and Z² or Z¹ and Z³,taken together, form -L- wherein L is —C(R^(a))₂, —C(R^(a))₂C(R^(a))₂—,—C(R^(a))₂C(R^(a))₂C(R^(a))₂—, or—C(R^(a))₂C(R^(a))₂C(R^(a))₂C(R^(a))₂—, wherein at least one of Z¹ andZ² or Z¹ and Z³, taken together, form -L-; Z⁴ is a bond, —CH₂—, or—CH₂CH₂—; Y¹ and Y² are each, independently, hydrogen, C₁₋₃alkyl orC₁₋₃haloalkyl; R¹ is phenyl substituted with one to three halogens; andeach R^(a) is, independently, hydrogen, halo, hydroxyl or C₁₋₄alkyl.2-108. (canceled)